CN1568373A - 介导rna干涉的小rna分子 - Google Patents

介导rna干涉的小rna分子 Download PDF

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CN1568373A
CN1568373A CNA018209009A CN01820900A CN1568373A CN 1568373 A CN1568373 A CN 1568373A CN A018209009 A CNA018209009 A CN A018209009A CN 01820900 A CN01820900 A CN 01820900A CN 1568373 A CN1568373 A CN 1568373A
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dsrna
sirna
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T·图舍尔
S·埃尔巴沙
W·兰德科尔
M·威尔姆
R·吕尔曼
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Europaisches Laboratorium fuer Molekularbiologie EMBL
Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
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Abstract

双链RNA(dsRNA)在多种生物中通过一种被称作RNA干涉(RNAi)的过程诱导序列特异的转录后基因沉默。利用果蝇体外系统,我们证明了19-23 nt短RNA片段是RNAi的序列特异性介体。短干涉RNA(siRNA)通过长dsRNA的RNase III样加工反应产生。化学合成的具有突出3’端的siRNA双链体在裂解液中介导高效的靶RNA切割,切割位点位于指导siRNA所覆盖的区域的中心附近。此外,我们还证明了dsRNA的加工方向决定是有义还是反义靶RNA能被产生的siRNP复合物切割。

Description

介导RNA干涉的小RNA分子
发明领域
本发明涉及介导靶特异的核酸修饰(如RNA干涉和/或DNA甲基化)所需的双链(ds)RNA分子的序列和结构特征。
发明背景
向线虫C.elegans中注射dsRNA导致在序列上与注射的dsRNA高度同源的基因特异性沉默(Fire等人,1998),在这一发现后,提出术语“RNA干涉”(RNAi)。之后也在昆虫、青蛙(Oelgeschlager等人,2000)和包括小鼠在内的其他动物(Svoboda等人,2000;Wianny和Zernicka-Goetz,2000)中观察到RNAi,在人类中也可能存在。RNAi与植物中共抑制和真菌中抑制(quelling)的转录后基因沉默(PTGS)机制密切相关(Catalanotto等人,2000;Cogoni和Macino,1999;Dalmay等人,2000;Ketting和Plasterk,2000;Mourrain等人,2000;Smardon等人,2000),RNAi机制的一些组分也是共抑制引起的转录后沉默所必需的(Catalanotto等人,2000;Dernburg等人,2000;Ketting和Plasterk,2000)。最近也对这一主题进行了综述(Bass,2000;Bosher和Labouesse,2000;Fire,1999;Plasterk和Ketting,2000;Sharp,1999;Sijen和kooter,2000),参见2000年3月2日发行的《植物分子生物学》第43卷的完整内容。
在植物中,除了PTGS之外,导入的转基因也能通过RNA指导的胞嘧啶DNA甲基化导致转录基因沉默(参见Wassenegger,2000)。植物中短至30bp的基因组靶标以RNA指导的方式甲基化(Pelissier,2000)。DNA甲基化在哺乳动物中也存在。
RNAi和共抑制的自然功能似乎是保护基因组免受移动遗传因子(例如在有活性时在宿主细胞中产生异常RNA或dsRNA的反转录转座子和病毒)的入侵(Jensen等人,1999;Ketting等人,1999;Ratcliff等人,1999;Tabara等人,1999)。特异性mRNA降解阻止转座子和病毒的复制,尽管某些病毒通过表达抑制PTGS的蛋白质能克服或阻止这一过程(Lucy等人,2000;Voinnet等人,2000)。
dsRNA只触发与dsRNA相同的区域内的同源RNA特异性降解(Zamore等人,2000)。dsRNA被加工为21-23nt RNA片段,靶RNA切割位点规则地间隔21-23nt。因此提示21-23nt片段是靶标识别的指导RNA(Zamore等人,2000)。在细胞裂解前用dsRNA转染黑尾果蝇(D.melanogaster Schneider)2细胞,在由该细胞制备的提取物中也检测到这种短RNA(Hammond等人,2000),然而,显示序列特异性核酸活性的级分也含有相当一部分残余dsRNA。观察到从加工的dsRNA中分离的21-23nt片段在一定程度上能介导特异性mRNA降解,这进一步支持了21-23nt片段在指导mRNA切割中的作用(Zamore等人,2000)。大小相似的RNA分子也在显示PTGS的植物组织中积累(Hamilton和Baulcombe,1999)。
在此,我们利用建立的果蝇(Drosophila)体外系统(Tuschl等人,1999;Zamore等人,2000)进一步研究RNAi机制。我们证实,当短21nt和22nt RNA与3’突出端碱基配对时,它们作为序列特异性mRNA降解的指导RNA。在该系统中短30bp dsRNA不能介导RNAi,因为它们不再加工为21nt和22nt RNA。此外,我们还确定了相对于21和22nt短干涉RNA(siRNA)的靶RNA切割位点,并且证实,dsRNA加工的方向决定了是有义还是反义靶RNA能被产生的siRNP内切核酸酶复合物切割。此外,siRNA也可能是转录调节的重要工具,例如通过引导DNA甲基化使哺乳动物基因沉默。
在人类体内细胞培养系统(HeLa细胞)中进行的其它实验表明,长度优选地为19-25个核苷酸的双链RNA分子具有RNAi活性。因此,与果蝇的结果不同,24和25nt长双链DNA分子对于RNAi也是有效的。
发明内容
本发明的目的在于提供能够介导靶特异性RNA干涉或其它靶特异性核酸修饰(如DNA甲基化)的新型试剂,这种试剂与现有试剂相比具有更高的效能和安全性。
一种分离的双链RNA分子提供了这一问题的解决方案,其中每条RNA链的长度为19-25,特别是19-23个核苷酸,其中该RNA分子能够介导靶特异性核酸修饰,特别是RNA干涉和/或DNA甲基化。优选地,至少一条链具有1-5个核苷酸、更优选地1-3个核苷酸、最优选地2个核苷酸的3’-突出端。另一条链可以是平端,或者具有可达6个核苷酸的3’突出端。如果dsRNA的两条链都正好是21或22nt,当两端都是平端时(0nt突出),也能观察到一定程度的RNA干涉。该RNA分子优选地是合成RNA分子,基本上不含细胞提取物,例如果蝇胚胎中存在的污染物。此外,该RNA分子优选地基本上不含任何非靶特异的污染物,特别是非靶特异的RNA分子,例如细胞提取物中存在的污染物。
此外,本发明还涉及分离的双链RNA分子的用途,其中每条RNA链的长度为19-25个核苷酸,用于介导哺乳动物细胞中,特别是人类细胞中靶特异的核酸修饰,特别是RNAi。
惊奇地发现,合成的短双链RNA分子,特别是具有突出3’端的,是序列特异的RNAi的介体,介导高效的靶-RNA切割,其中切割位点靠近指导短RNA覆盖的区域的中心。
优选地,RNA分子的每条链的长度为20-22个核苷酸(或者在哺乳动物细胞中为20-25个核苷酸),其中每条链的长度可以相同或者不同。3’-突出端的长度优选地达到1-3个核苷酸,其中每条链的突出端长度可以相同或者不同。RNA-链优选地含有3’-羟基。5’-端优选地含有磷酸、二磷酸、三磷酸或羟基。最有效的dsRNA由配对的两条21nt链组成,使得dsRNA的两端存在1-3nt,特别是2nt的3’突出端。
siRNA指导的靶RNA切割反应是高度序列特异的。然而,不是siRNA的所有位点对于靶标识别都有相同的作用。在siRNA双链体中心的错配最为关键,基本上消除靶RNA切割。相反,与单链靶RNA互补的siRNA链的3’核苷酸(例如位点21)对于靶标识别的特异性就没有作用。此外,与靶RNA具有相同极性的siRNA链的未配对2nt 3’突出端的序列对于靶RNA切割并不重要,因为只有反义siRNA链才能指导靶标识别。因此,从单链突出核苷酸起,只有反义siRNA的倒数第二个位点(例如位点20)需要匹配靶向的有义mRNA。
令人吃惊的是,本发明的双链RNA分子在血清或用于细胞培养的生长培养基中显示很高的体内稳定性。为了进一步提高稳定性,可以稳定3’突出端使其免受降解,例如,可以加以选择,使其由嘌呤核苷酸,特别是腺苷或鸟苷核苷酸组成。此外,嘧啶核苷酸置换为修饰类似物,例如尿苷2nt 3’突出端置换为2’-脱氧胸苷可以耐受,不影响RNA干涉的效率。不含2’羟基可以显著提高突出端在组织培养基中的核酸酶抗性。
在本发明的一个特别优选的实施方案中,RNA分子可含有至少一种修饰的核苷酸类似物。该核苷酸类似物可位于基本上不影响靶特异性活性(如RNAi介导活性)的位置上,例如位于双链RNA分子的5’端和/或3’端区。特别是,掺入修饰的核苷酸类似物可以稳定突出端。
优选的核苷酸类似物选自糖修饰或骨架修饰的核糖核苷酸。然而应当指出,核碱基修饰的核糖核苷酸也是合适的,即核糖核苷酸,其含有非天然存在的核碱基代替天然存在的核碱基,如5-位修饰的尿苷或胞苷,例如5-(2-氨基)丙基尿苷、5-溴尿苷;8-位修饰的腺苷和鸟苷,例如8-溴鸟苷;去氮杂核苷酸,例如7-去氮杂-腺苷;O-和N-烷基化核苷酸,例如N6-甲基腺苷。在优选的糖修饰的核糖核苷酸中,2’OH-基被替换为选自H、OR、R、卤素、SH、SR、NH2、NHR、NR2或CN的基团,其中R是C1-C6烷基、烯基或炔基,卤素是F、Cl、Br或I。在优选的骨架修饰的核糖核苷酸中,与相邻核糖核苷酸连接的磷酯基团被替换为一个修饰的基团,例如硫代磷酸酯(phosphothioate)基。应当指出,上述修饰可以组合。
为了介导靶特异的RNAi和/或DNA甲基化,本发明的双链RNA分子的序列必须与核酸靶分子具有足够的同一性。优选地,该序列与RNA分子双链部分中的希望的靶分子至少50%,特别是至少70%相同。在RNA分子的双链部分中,同一性更优选地至少85%,最优选地100%。一种双链RNA分子与预定的核酸靶分子(如mRNA靶分子)的同一性可以如下确定:
I = - n L × 100
其中I是百分同一性,n是dsRNA的双链部分中与靶标相同的核苷酸数,L是dsRNA的双链部分与靶标的序列重叠的长度。
此外,也可包括3’突出端,特别是长度为1-3个核苷酸的突出端,确定双链RNA分子与靶序列的同一性。在此情况中,与靶序列的序列同一性优选地至少50%,更优选地至少70%,最优选地至少85%。例如,来自3’突出端的核苷酸和来自双链5’和/或3’端的可达2个核苷酸可以修饰,而活性没有显著丧失。
本发明的双链RNA分子可以用包括下列步骤的方法制备:
(a)合成两条RNA链,每条长度为19-25个,例如19-23个核苷酸,其中该RNA链能形成双链RNA分子,优选地至少一条链具有1-5个核苷酸的3’-突出端,
(b)在一定条件下结合合成的RNA链,其中形成一种双链RNA分子,它能介导靶特异的核酸修饰,特别是RNA干涉和/或DNA甲基化。
合成RNA分子的方法在本领域中众所周知。在本文中,具体是指如Verma和Eckstein(1998)所述的化学合成法。
由合成的DNA模板或者由分离自重组细菌的DNA质粒进行酶转录也能制备单链RNA。一般使用噬菌体RNA聚合酶,如T7、T3或SP6 RNA聚合酶(Milligan和Uhlenbeck(1989))。
本发明的另一个方面涉及一种介导细胞或生物中的靶特异性核酸修饰(特别是RNA干涉和/或DNA甲基化)的方法,包括下列步骤:
(a)在可以发生靶特异性核酸修饰的条件下使细胞或生物接触本发明的双链RNA分子,和
(b)将双链RNA完成的靶特异性核酸修饰导向含有基本对应于双链RNA的序列部分的靶核酸。
优选地,接触步骤(a)包括将双链RNA分子导入靶细胞中,例如分离的靶细胞,例如细胞培养物,单细胞微生物,或多细胞生物内的一个靶细胞或多个靶细胞。更优选地,该导入步骤包括载体介导的输送,例如利用脂质体载体或通过注射。
本发明的方法可用来确定细胞或生物中一种基因的功能,乃至调节细胞或生物中一种基因的功能,它能够介导RNA干涉。细胞优选地是真核细胞或细胞系,例如植物细胞或动物细胞,如哺乳动物细胞,例如胚细胞、多能干细胞、肿瘤细胞,例如畸胎癌细胞或病毒感染的细胞。生物优选地是真核生物,例如植物或动物,如哺乳动物,特别是人类。
本发明的RNA分子针对的靶基因可能与病理状态有关。例如,该基因可能是病原体相关基因,例如病毒基因,肿瘤相关基因,或自身免疫病相关基因。靶基因也可能是在重组细胞或遗传改变的生物中表达的异源基因。通过确定或调节,特别是抑制这种基因的功能,可能获得在农业领域或在医学或兽医学领域有价值的信息和治疗效益。
dsRNA通常作为药用组合物施用。可用已知方法进行施用,其中向体外或体内的希望的靶细胞中导入核酸。常用的基因转移技术包括磷酸钙、DEAE-葡聚糖、电穿孔、显微注射和病毒法(Graham,F.L.和van der Eb,A.J.(1973)Virol.52,456;McCutchan,J.H.和Pagano,J.S.(1968),J.Natl.Cancer Inst.41,351;Chu,G.等人(1987),Nucl.AcidsRes.15,1311;Fraley,R.等人(1980),J.Biol.Chem.255,10431;Capecchi,M.R.(1980),Cell 22,479)。向细胞中导入DNA的一种最新技术是使用阳离子脂质体(Felgner,P.L.等人(1987),Proc.Natl.Acad.Sci USA 84,7413)。市售的阳离子脂制剂有,例如Tfx 50(Promega)或Lipofectamin2000(Life Technologies)。
因此,本发明也涉及一种药用组合物,其含有至少一种上述双链RNA分子作为活性剂和一种药用载体。该组合物可在人类医学或兽医学中用于诊断和治疗用途。
为用于诊断或治疗,该组合物可以是溶液形式,例如可注射溶液,乳膏、软膏、片剂、悬液等。该组合物可以用任何适当的方法施用,例如注射、口服、局部、鼻、直肠给药等。载体可以是任何合适的药用载体。优选地使用能提高RNA分子进入靶细胞的效能的载体。这类载体的合适的例子有脂质体,特别是阳离子脂质体。更加优选的施用方法是注射。
RNAi法的另一个优选用途是对真核细胞或真核非人类生物的功能分析,优选的是哺乳动物细胞或生物,最优选的是人类细胞,例如细胞系,如HeLa或293,或啮齿动物,例如大鼠或小鼠。通过用与预定靶基因同源的适当双链RNA分子或编码适当双链RNA分子的DNA分子转染,能够在靶细胞,例如细胞培养物或靶生物中获得特定敲除表型。惊奇地发现,存在短双链RNA分子不会引起宿主细胞或宿主生物的干扰素反应。
因此,本发明的另一个主题是显示靶基因特异的敲除表型的真核细胞或真核非人类生物,其中包含至少一种内源靶基因的表达至少部分缺陷,其中用能够抑制至少一种内源靶基因表达的至少一种双链RNA分子或用编码能够抑制至少一种内源靶基因表达的至少一种双链RNA分子的DNA转染该细胞或生物。应当指出,由于RNAi的特异性,本发明能够靶特异性地敲除几种不同的内源基因。
细胞或非人类生物,特别是人类细胞或非人类哺乳动物的基因特异性敲除表型可以在分析方法中使用,例如复杂生理过程的功能和/或表型分析,如基因表达谱和/或蛋白质组的分析。例如,人们可以在培养的细胞中制备人类基因的敲除表型,假定它们是备选剪接过程的调节剂。这类基因具体包括SR剪接因子家族的成员,如ASF/SF2、SC35、SRp20、SRp40或SRp55。另外也能分析SR蛋白对预定的其它剪接基因(如CD44)的mRNA谱的影响。优选地利用基于寡核苷酸的芯片通过高通量方法进行这种分析。
利用基于RNAi的敲除技术,可以抑制靶细胞或靶生物中内源靶基因的表达。内源基因可以用编码靶蛋白或靶蛋白变体或突变形式的外源靶核酸(例如基因或cDNA)补充,外源核酸任选地可与编码可检测肽或多肽(例如亲和标记物,特别是多亲和标记物)的另一种核酸序列融合。靶基因的变体或突变形式不同于内源靶基因,因为由于单个或多个氨基酸的置换、插入和/或缺失,它们编码在氨基酸水平上与内源基因产物不同的基因产物。变体或突变形式可具有与内源靶基因相同的生物活性。另一方面,变异或突变的靶基因也可能具有不同于内源靶基因的生物活性,例如活性部分缺失、活性完全缺失、活性提高等。
这种互补可以如下完成:共表达外源核酸编码的多肽,例如含有靶蛋白和亲和标记物的融合蛋白和用于敲除靶细胞中内源基因的双链RNA分子。这种共表达可以利用一种合适的表达载体或者利用表达载体的组合实现,该载体表达外源核酸编码的多肽(例如标记物修饰的靶蛋白)和双链RNA分子。在靶细胞中从头合成的蛋白质和蛋白质复合物将含有外源基因产物,例如修饰的融合蛋白。为了避免RNAi双链分子抑制外源基因产物表达,编码外源核酸的核苷酸序列在与双链RNA分子同源的序列部分中可能在DNA水平上改变(在氨基酸水平上引起或不引起突变)。此外,也可以用来自其它种(例如来自小鼠)的相应核苷酸序列补充内源靶基因。
本发明的细胞或生物的优选用途是对基因表达谱和/或蛋白质组的分析。在一个特别优选的实施方案中,对一种或几种靶蛋白的变体或突变形式进行分析,其中利用如上所述的外源靶核酸将这种变体或突变形式重新导入细胞或生物中。内源基因敲除与利用突变的(例如部分缺失的)外源靶标拯救相组合比使用敲除细胞具有优势。而且,这种方法还特别适用于鉴定靶蛋白的功能域。在另一个优选实施方案中,例如对至少两种细胞或生物的基因表达谱和/或蛋白质组和/或表型特征进行比较。这些生物选自:
(i)一种对照细胞或对照生物,没有靶基因抑制,
(ii)一种细胞或生物,具有靶基因抑制,和
(iii)一种细胞或生物,具有靶基因抑制加外源靶核酸的靶基因互补。
本发明的方法和细胞也适用于鉴定和/或表征药理试剂的方法,例如从一组待测物质中鉴定新的药理试剂,和/或表征已知药理试剂的作用机制和/或副作用。
因此,本发明也涉及一种鉴定和/或表征作用于至少一种靶蛋白的药理试剂的系统,其包括:
(a)一种真核细胞或真核非人类生物,它能表达至少一种编码该靶蛋白的内源靶基因,
(b)至少一种双链RNA分子,它能抑制所述至少一种内源靶基因的表达,和
(c)一种待测物质或一组待测物质,其中将要鉴定和/或表征这种待测物质或组合的药理学性质。
如上所述的系统优选地还包括:
(d)至少一种编码靶蛋白或其变体或其突变形式的外源靶核酸,其中该外源靶核酸在核酸水平上不同于内源靶基因,因此与内源靶基因的表达相比,外源靶核酸的表达受双链RNA分子的抑制大大减少。
此外,RNA敲除互补法还可用于制备目的,例如从真核细胞,特别是哺乳动物细胞,更具体地是从人类细胞中亲和纯化蛋白质或蛋白质复合物。在本发明的实施方案中,外源靶核酸优选地编码与亲和标记物融合的靶蛋白。
该制备方法可以用于高分子量蛋白质复合物的纯化,其分子量优选地≥150kD,更优选地≥500kD,任选地可含有核酸,如RNA。具体例子有:由U4/U6 snRNP颗粒的20kD、60kD和90kD蛋白质组成的异源三聚蛋白质复合物,由分子量14、49、120、145和155kD的5种蛋白质组成的来自17S U2 snRNP的剪接因子SF3b,含有U4、U5和U6 snRNA分子和大约30种蛋白质的25S U4/U6/U5三-snRNP颗粒,其分子量约为1.7MD。
该方法适用于哺乳动物细胞,特别是人类细胞的功能蛋白质组分析。
下面,通过下列附图和实施例更详细地说明本发明。
附图说明
图1:短至38bp的双链RNA能介导RNAi。
(A)用于靶向Pp-Luc mRNA的dsRNA的图示。制备覆盖29-504bp的三个系列的平端dsRNA。标出了dsRNA有义链的第一个核苷酸相对于Pp-Luc mRNA起始密码子的位置(p1)。(B)RNA干涉测定(Tuschl等人,1999)。靶Pp-Luc与对照Rr-luc活性之比用缓冲液对照(黑条)标准化。dsRNA(5nM)在果蝇裂解液中25℃预温育15分钟,之后加入7-甲基-鸟苷-加帽的Pp-luc和Rr-luc mRNA(~50pM)。继续温育1小时,然后通过双萤光素酶测定(Promega)分析。数据是至少4次独立实验的平均值±标准差。
图2:29bp dsRNA不再加工为21-23nt片段。在果蝇裂解液中内部32P-标记的dsRNA(5nM)加工后形成21-23mer的时程。标出了dsRNA的长度和来源。RNA分子量标记(M)加于左道中,并标出片段大小。时间为0时的双条带是由于dsRNA的不完全变性。
图3:短dsRNA只切割mRNA靶标一次。
(A)10nM帽32P-标记的有义或反义RNA与p133系列的10nMdsRNA在果蝇裂解液中温育1小时,产生的稳定5’切割产物的变性凝胶电泳。帽标记的靶RNA部分核酸酶T1消化并部分碱性水解(OH)产生长度标记物。dsRNA针对的区域在两端表示为黑条。显示了111bp长dsRNA的主要切割位点之间的20-23nt间隔。水平箭头表明不是由于RNAi引起的非特异性切割。(B)有义和反义靶RNA上切割位点的位置。加帽177nt有义和180nt反义靶RNA的序列以反平行方向表示,使得互补序列彼此相对。不同dsRNA针对的区域用位于有义与反义靶序列之间的不同颜色的线条标出。切割位点用环形标出:大环为强切割,小环为弱切割。32P-放射性标记的磷酸基以星号标记。
图4:21和22nt RNA片段通过RNase III样机制产生。
(A)dsRNA加工后~21nt RNA的序列。定向克隆dsRNA加工产生的~21nt RNA片段并测序。来源于dsRNA有义链的寡核糖核苷酸用蓝线表示,来源于反义链的用红线表示。如果在多个克隆中含有相同的序列,则使用粗线,右侧的数字是指频率。dsRNA介导的靶RNA切割位点用橙色环形表示,大环表示强切割,小环表示弱切割(参见图3B)。有义链之上的环形表示有义靶标内的切割位点,dsRNA下面的环形表示反义靶标中的切割位点。在来源于dsRNA 3’端的~21nt片段中鉴定出高达5个其它核苷酸。这些核苷酸主要是C、G或A残基的随机组合,最可能是在dsRNA组成链的T7转录过程中以非模板方式添加的。(B)~21nt RNA的核苷酸组成的二维TLC分析。~21nt RNA如下产生:内部放射性标记的504bp Pp-luc dsRNA在果蝇裂解液中温育,凝胶纯化,然后用核酸酶P1(最上一行)或核糖核酸酶T2(最下一行)消化为单核苷酸。在存在指定α-32P核苷三磷酸之一的情况下,通过转录在内部放射性标记dsRNA。放射性通过磷成像检测。核苷5’-一磷酸、核苷3’-一磷酸、核苷5’,3’-二磷酸和无机磷酸盐分别表示为pN、Np、pNp和Pi。黑色环形表示非放射性载体核苷酸的UV吸收点。用T4多核苷酸激酶和γ-32P-ATP使核苷3’-一磷酸5’磷酸化,制备放射性标记标准,通过与该标准共迁移鉴定3’,5’-二磷酸(红色环形)。
图5:合成的21和22nt RNA介导靶RNA切割。
(A)对照52bp dsRNA和合成21和22nt dsRNA的图示。21和22nt短干涉RNA(siRNA)的有义链用蓝色表示,反义链用红色表示。除了双链体5的22nt反义链之外,siRNA的序列来源于52和111bpdsRNA的克隆片段(图4A)。双链体6和7中的siRNA是111bp dsRNA加工反应所特有的。在双链体1和3的合成反义链序列中存在两个3’突出核苷酸,以绿色表示。对照52bp dsRNA的两条链都通过体外转录制备,一部分转录物可能包括非模板3’核苷酸添加。siRNA双链体指导的靶RNA切割位点表示为橙色环形(见图4A的图例),如图5B所示确定。(B)有义和反义靶RNA上切割位点的位置。靶RNA序列如图3B所示。对照52bp dsRNA(10nM)或21和22nt RNA双链体1-7(100nM)与靶RNA一起在果蝇裂解液中25℃温育2.5小时。稳定的5’切割产物在凝胶上分析。切割位点在图5A中标出。52bp dsRNA针对的区域或有义或反义链在凝胶旁边以黑线标出。切割位点全都位于与dsRNA相同的区域内。为了精确确定反义链的切割位点,使用较低百分比的凝胶。
图6:短dsRNA上的长3’突出端抑制RNAi。
(A)52bp dsRNA构建体的图示。有义和反义链的3’延伸分别以蓝色和红色表示。靶RNA上的切割位点用类似于图4A的橙色环形表示,如图6B所示确定。(B)有义和反义靶RNA上切割位点的位置。靶RNA序列如图3B所示。dsRNA(10nM)与靶RNA一起在果蝇裂解液中25℃温育2.5小时。稳定的5’酶切产物在凝胶上分析。主要切割位点用水平箭头标出,也在图6A中表示。52bp dsRNA针对的区域在凝胶两侧以黑线表示。
图7:建议的RNAi模型。
预测RNAi从dsRNA(有义链为黑色,反义链为红色)主要加工为21和22nt短干扰RNA(siRNA)开始。如果dsRNA上存在短的突出3’核苷酸,它们可能有利于短dsRNA的加工。有待表征的dsRNA加工蛋白用绿色和蓝色椭圆形表示,在dsRNA上以不对称方式装配。在我们的模型中,假定的蓝色蛋白或蛋白域与siRNA链以3’-5’方向结合,而假定的绿色蛋白或蛋白域总是结合相对的siRNA链。这些蛋白质或亚组与siRNA双链体结合,并且保持其方向,这取决于dsRNA加工反应的方向。只有与蓝色蛋白结合的siRNA序列才能指导靶RNA切割。内切核酸酶复合物被称为小干扰核糖核蛋白复合物或siRNP。在此我们假定,切割dsRNA的内切核酸酶也可切割靶RNA,这可能是通过暂时置换不用于靶标识别的被动siRNA链。靶RNA然后在序列互补指导siRNA识别的区域中心切割。
图8:报道构建体和siRNA双链体。
(a)显示来自质粒pGL2-对照、pGL-3-对照和pRL-TK(Promega)的萤火虫(Pp-luc)和海肾(Rr-luc)萤光素酶报道基因区。显示SV40调节元件、HSV胸苷激酶启动子和两个内含子(线)。GL3萤光素酶的序列与GL2有95%相同,但RL与这两者完全无关。在转染的哺乳动物细胞中由pGL2表达萤光素酶比由pGL3表达大约低10倍。SiRNA双链体针对的区域在萤光素酶基因编码区下用黑线表示。(b)显示针对GL2、GL3和RL萤光素酶的siRNA双链体的有义(上)和反义(下)序列。GL2和GL3 siRNA双链体只有3个单核苷酸置换(以灰色框出)的不同。合成含有反向GL2序列的双链体invGL2作为非特异对照。2’-脱氧胸苷的2nt 3’突出端表示为TT;uGL2类似于GL2 siRNA,但是含有核糖尿苷3’突出端。
图9:siRNA双链体引起的RNA干涉。
靶与对照萤光素酶之比用缓冲液对照(bu,黑线)标准化;灰线表示Photinus pyralis(Pp-luc)GL2或GL3萤光素酶与Renillareniformis(Rr-luc)RL萤光素酶之比(左轴),白线表示RL与GL2或GL3之比(右轴)。图a、c、e、g、i描述用pGL2-对照和pRL-TK报道质粒的组合进行的实验,图b、d、f、h、j使用pGL3-对照和pRL-TK报道质粒。用于干涉实验的细胞系在每张图的顶部标出。在标准化之前及在所检测的不同细胞系之间,缓冲液对照(bu)的Pp-luc/Rr-luc比分别为:pGL2/pRL:0.5-10,pGL3/pRL:0.03-1。作图的数据是三次独立实验的平均值±S.D.。
图10:21nt siRNA、50bp和500bp dsRNA对HeLa细胞中萤光素酶表达的影响。
长dsRNA的确切长度在线条下面标出。图a、c、e描述用pGL2-对照和pRL-TK报道质粒进行的实验,图b、d、f使用pGL3-对照和pRL-TK报道质粒。数据是两次独立实验的平均值±S.D.。(a),(b)绝对Pp-luc表达,以任意发光单位作图。(c),(d)Rr-luc表达,以任意发光单位作图。(e),(f)标准化靶标与对照萤光素酶之比。siRNA双链体的萤光素酶活性之比用缓冲液对照(bu,黑线)标准化;50或500bpdsRNA的发光比分别用来自人源化GFP(hG,黑线)的50或500bpdsRNA的发光比标准化。应当指出,靶向GL2和GL3的49与484bpdsRNA之间的总体序列差异不足以使GL2与GL3靶标之间具有特异性(49bp片段中有43nt连续同一性,484bp片段中有239nt最长连续同一性)。
图11:21-nt siRNA双链体3’突出端的变异。
(A)实验策略概述。显示了加帽的和聚腺苷酸化的有义靶mRNA,并且显示了有义和反义siRNA的相对位置。根据8个不同的反义链,制备了8个系列的双链体。siRNA序列和突出核苷酸的数量以1nt的幅度改变。(B)在5nM平端dsRNA存在下,用黑尾果蝇胚胎裂解液中的对照萤光素酶(Renilla reniformis,Rr-luc)标准化的靶萤光素酶(Photinus pyralis,Pp-luc)的相对发光。在dsRNA存在下测定的发光比用缓冲液对照(bu,黑线)获得的发光比标准化。标准化的发光比小于1表明有特异性干涉。(C-J)8个系列21-nt siRNA双链体的标准化干涉比。siRNA双链体的序列在条图之上显示。每张图显示由特定反义指导siRNA和5种不同的有义siRNA形成的一组双链体的干涉比。突出核苷酸的数量(3’突出端,正数;5’突出端,负数)在x轴上标出。数据点是至少3次独立实验的平均值,误差条代表标准差。
图12:siRNA双链体有义链长度的变化。
(A)实验图示。3条21-nt反义链与8个有义siRNA配对。这些siRNA的3’端长度改变。反义siRNA的3’突出端为1-nt(B)、2-nt(C)或3-nt(D),而每个系列的有义siRNA突出端不同。标出了siRNA双链体的序列和相应的干涉比。
图13:保留2-nt 3’突出端的siRNA双链体的长度变化。
(A)实验图示。21-nt siRNA双链体在序列上与图11H或图12C所示相同。这些siRNA双链体延伸到有义siRNA的3’端(B)或有义siRNA的5’端(C)。标出了siRNA双链体的序列和各自的干涉比。
图14:siRNA核糖残基的2’-羟基的置换。
将siRNA双链体链中的2’-羟基(OH)置换为2’-脱氧(d)或2’-O-甲基(Me)。3’端的2-nt和4-nt 2’-脱氧置换分别表示为2-nt d和4-nt d。尿苷残基置换为2’-脱氧胸苷。
图15:具有2-nt 3’突出端的21-nt siRNA双链体对有义和反义靶RNA切割的作图。
(A)32P(星号)帽标记的有义和反义靶RNA和siRNA双链体的图示。有义和反义靶RNA切割的位置分别在siRNA双链体之上和之下用三角形标出。(B)靶RNA切割位点的作图。10nM靶标与100nMsiRNA双链体在黑尾果蝇胚胎裂解液中温育2小时后,在测序凝胶上分析5’帽标记的底物和5’切割产物。长度标记物通过靶RNA的部分RNase T1消化(T1)和部分碱性水解(OH-)产生。图左侧的粗线表示与靶标同向的siRNA链1和5所覆盖的区域。
图16:指导siRNA的5’端决定靶RNA切割位置。
(A,B)实验策略图示。所有siRNA双链体中的反义siRNA相同,但是通过改变3’端有义链有18-25nt的不同(A),或者通过改变5’端有18-23nt的不同(B)。有义和反义靶RNA切割位置分别在siRNA双链体之上和之下用三角形标出。(C,D)利用帽标记的有义(上图)或反义(下图)靶RNA对靶RNA切割的分析。只显示帽标记的5’切割产物。标出了siRNA双链体的序列,有义siRNA链的长度在图上部标出。图(C)中用破折号标记的对照道显示在不含siRNA情况下温育的靶RNA。标记物如图15所述。(D)下图中的箭头指出相差1nt的靶RNA切割位点。
图17:siRNA双链体3’突出端的序列变异。
2-nt 3’突出端(NN,灰色)如图所示在序列和组成上有变化(T,2’-脱氧胸苷,dG,2’-脱氧鸟苷,星号,野生型siRNA双链体)。标准化的干涉比如图11所述确定。野生型序列与图14所示相同。
图18:靶标识别的序列特异性。
显示了错配siRNA双链体的序列,修饰的序列片段或单核苷酸以灰色在下面列出。参照双链体(ref)和siRNA双链体1-7含有2’-脱氧胸苷2-nt突出端。胸苷修饰的参照双链体的沉默率与野生型序列相当(图17)。标准化的干涉比如图11所述确定。
图19:保留2-nt 3’突出端的siRNA双链体的长度变化。
siRNA双链体延伸至有义siRNA的3’侧(A)或有义siRNA的5’侧(B)。标出了siRNA双链体的序列和各自的干涉比。对于HeLa SS6细胞,靶向GL2萤光素酶的siRNA双链体(0.84μg)用pGL2-对照和pRL-TK质粒一起转染。为了进行对比,标出了在黑尾果蝇裂解液中检测到的siRNA双链体的体外RNAi活性。
实施例1
合成小RNA介导的RNA干涉
1.1.实验方法
1.1.1体外RNAi
体外RNAi和裂解液的制备如前所述进行(Tuschl等人,1999;Zamore等人,2000)。使用新溶解的肌酸激酶(Roche)对于最佳ATP再生至关重要。在25℃下,经过15分钟的延长预温育时间,用5nM的dsRNA浓度进行RNAi翻译测定(图1),之后加入体外转录、加帽并且聚腺苷酸化的Pp-luc和Rr-luc报道mRNA。继续温育1小时,利用双萤光素酶测定(Promega)和单光3010C发光计(PharMingen)分析Pp-luc和Rr-luc蛋白的相对含量。
1.1.2RNA合成
由携带T7或SP6启动子序列的PCR模板体外转录RNA使用标准方法,例如参见(Tuschl等人,1998)。合成的RNA用Expedite RNA亚磷酰胺(Proligo)制备。3’接头寡核苷酸用二甲氧基三苯甲基-1,4-苯二甲醇-琥珀酰-氨丙基-CPG合成。寡核糖核苷酸在3ml 32%氨/乙醇(3/1)中55℃去保护4小时(Expedite RNA)或55℃ 16小时(3’和5’接头DNA/RNA嵌合寡核苷酸),然后脱硅烷化,如前所述凝胶纯化(Tuschl等人,1993)。包含长3’突出端的dsRNA制品的RNA转录物由有义方向含有T7启动子、反义方向含有SP6启动子的PCR模板产生。用GCG TAATACGACTCACTATAGAACAATTGCTTTTACAG(下划线标出T7启动子)作为5’引物,用ATTTAGGTGACACTATAGGCATAAAGAATTGAAGA(下划线标出SP6启动子)作为3’引物,用线性化Pp-luc质粒(pGEM-luc序列)(Tuschl等人,1999)作为模板,PCR扩增有义和反义靶RNA的转录模板;T7转录的有义RNA为177nt长,在相对于起始密码子的位点113-273之间含有Pp-luc序列,之后在3’端是SP6启动子序列的17nt互补序列。通过只含单启动子序列的两种不同PCR产物的转录,制备用于形成平端dsRNA的转录物。
利用苯酚/氯仿抽提进行dsRNA退火。等摩尔浓度的有义和反义RNA(50nM-10μM,取决于长度和含量)在0.3M NaOAc(pH6)中90℃温育30秒,然后在室温下用等体积的苯酚/氯仿抽提,之后用氯仿抽提除去残余的苯酚。加入2.5-3倍体积的乙醇沉淀产生的dsRNA。沉淀溶解于裂解缓冲液(100mM KCl,30mM HEPES-KOH,pH7.4,2mM Mg(OAc)2)中,在1×TAE缓冲液中通过标准琼脂糖凝胶电泳证实dsRNA的质量。具有17nt和20nt 3’突出端的52bp dsRNA(图6)在95℃下温育1分钟退火,迅速冷却到70℃,然后在3个小时内缓慢冷却到室温(50μl退火反应,1μM标准浓度,300mM NaCl,10mMTris-HCl,pH7.5)。然后用苯酚/氯仿抽提dsRNA,乙醇沉淀,溶解于裂解缓冲液中。
用1mM ATP、CTP、GTP、0.1或0.2mM UTP和0.2-0.3μM32P-UTP(3000 Ci/mmol),或者用是放射性标记核苷三磷酸的各自的比例而不是UTP的比例转录用于制备dsRNA的内部32P-放射性标记的RNA(图2和图4)。靶RNA的帽子的标记如前所述进行。靶RNA在帽子标记后凝胶纯化。
1.1.3切割位点作图
标准RNAi反应如下进行:预温育10nM dsRNA 15分钟,随后加入10nM帽子标记的靶RNA。再温育2小时(图2A)或2.5小时(图5B和6B)后,通过蛋白酶K处理终止反应(Tuschl等人,1999)。然后在8%或10%测序凝胶上分析样品。21和22nt合成RNA双链体以100nM终浓度使用(图5B)。
1.1.4~21nt RNA的克隆
在不含靶RNA的情况下,在果蝇裂解液中温育放射性标记的dsRNA,产生21nt RNA(200μl反应,1小时温育,50nM dsP111,或100nM dsP52或dsP39)。然后用蛋白酶K处理反应混合物(Tuschl等人,1999),在变性15%聚丙烯酰胺凝胶上分离dsRNA加工产物。切下大小范围至少为18-24nt的条带,洗脱到0.3M NaCl中,在4℃下在硅化试管中过夜。通过乙醇沉淀回收RNA,去磷酸化(30μl反应,30分钟,50℃,10U碱性磷酸酶,Roche)。通过苯酚/氯仿抽提终止反应,乙醇沉淀RNA。3’接头寡核苷酸(pUUUaaccgcatccttctcx:大写,RNA;小写,DNA;p,磷酸;x,4-羟甲基苄基)与去磷酸化的~21ntRNA连接(20μl反应,30分钟,37℃,5μM 3’接头,50mM Tris-HCl,pH7.6,10mM MgCl2,0.2mM ATP,0.1mg/ml乙酰BSA,15%DMSO,25U T4 RNA连接酶,Amersham-Pharmacia)(Pan和Uhlenbeck,1992)。加入等体积的8M尿素/50mM EDTA终止混合物终止连接反应,直接加样到15%凝胶上。连接产率超过50%。从凝胶中回收连接产物,并5’-磷酸化(20μl反应,30分钟,37℃,2mM ATP,5U T4多核苷酸激酶,NEB)。通过苯酚/氯仿抽提终止磷酸化反应,乙醇沉淀回收RNA。然后,5’接头(tactaatacgactcactAAA:大写,RNA;小写,DNA)如上所述与磷酸化连接产物连接。凝胶纯化新的连接产物,在用作载体的反转录引物(GACTAGCTG GAATTCAAGGATGCGGTTAAA,下划线,EcoRI位点)存在下,从凝胶切片中洗脱。反转录(15μl反应,30分钟,42℃,150U Superscript II反转录酶,Life Technologies)之后用CAGCCAACG GAATTCATACGACTCACTAAA(下划线,EcoRI位点)作为5’引物和3’RT引物进行PCR。经苯酚/氯仿抽提和乙醇沉淀纯化PCR产物。然后用EcoRI(NEB)消化PCR产物,用T4 DNA连接酶(高浓度,NEB)连环化。在低熔点琼脂糖凝胶上分离大小为200-800bp的多联体,利用标准的熔化和苯酚抽提法从凝胶中回收,乙醇沉淀。在标准条件下与Taq聚合酶72℃温育15分钟,补平未配对的末端,用TOTO TA克隆试剂盒(Invitrogen)将DNA产物与pCR2.1-TOPO载体直接连接。利用PCR和M13-20和M13反向测序引物筛选集落。直接对PCR产物进行委托测序(Sequence Laboratories GttingenGmbH,德国)。每个克隆平均获得4-5条21mer序列。
1.1.5 2D-TLC分析
放射性标记、凝胶纯化的siRNA的核酸酶P1消化和2D-TLC如(Zamore等人,2000)所述进行。采用2μg/μl载体tRNA和30U核糖核酸酶T2(Life Technologies),在10mM乙酸铵(pH4.5)中以10μl反应体积进行核酸酶T2消化,50℃ 3小时。非放射性标准的迁移通过UV影像法测定。T2消化产物与使用γ-32P-ATP和T4多核苷酸激酶经商品核苷3’-一磷酸的5’-32P-磷酸化制备的标准共迁移,由此证实核苷-3’,5’-二磷酸的身份(数据未显示)。
1.2结果与讨论
1.2.1dsRNA加工为21和22nt RNA片段的长度要求
由黑尾果蝇合胞体胚胎制备的裂解液概述了体外RNAi,提供了一个用于生化分析RNAi机制的新型工具(Tuschl等人,1999;Zamore等人,2000)。RNAi对dsRNA的长度要求的体外和体内分析揭示,在降解靶mRNA上,短dsRNA(<150bp)比长dsRNA效率更低(Caplen等人,2000;Hammond等人,2000;Ngo等人,1998;Tuschl等人,1999)。mRNA降解效率降低的原因尚不清楚。因此我们在最佳条件下,在果蝇裂解液中,检查了靶RNA降解对dsRNA的精确长度要求(Zamore等人,2000)。合成了几个系列的dsRNA,它们针对萤火虫萤光素酶(Pp-luc)报道RNA。用双萤光素酶测定(Tuschl等人,1999)监测靶RNA表达的特异性抑制(图1A和1B)。对于短至38bp的dsRNA,我们检测到靶RNA表达的特异性抑制,但是29-36bp的dsRNA在该方法中无效。此作用与靶位点无关,与dsRNA长度(即长dsRNA)相关的Pp-luc mRNA表达的抑制程度比短dsRNA更有效。
曾经提出,dsRNA加工产生的21-23nt RNA片段是RNA干涉和共抑制的介体(Hamilton和Baulcombe,1999;Hammond等人,2000;Zamore等人,2000)。因此我们分析了大小为501-29bp的dsRNA亚组的21-23nt片段形成率。对于39-501bp长dsRNA,容易地检测到果蝇裂解液中21-23nt片段的形成(图2),但是29bp dsRNA明显延迟。这一发现与21-23nt片段在指导mRNA切割上的作用一致,为RNAi缺乏30bp dsRNA提供了一种解释。21-23mer形成的长度信赖性可能反映了一种生物学相关的控制机制,其通过规则细胞RNA的分子内碱基配对的短结构阻止不希望的RNAi激活。
1.2.2 39bp dsRNA介导靶RNA在单个位点处的切割
向果蝇裂解液中加入dsRNA和5’-加帽的靶RNA导致靶RNA的序列特异性降解(Tuschl等人,1999)。靶mRNA只在与dsRNA相同的区域内切割,许多靶切割位点相隔21-23nt(Zamore等人,2000)。因此,特定dsRNA的切割位点数预计大约对应于dsRNA的长度除以21。我们对在帽子处5’放射性标记的有义和反义靶RNA上的靶切割位点作图(Zamore等人,2000)(图3A和3B)。在测序凝胶上分离稳定的5’切割产物,通过与来自靶RNA的部分RNase T1和碱性水解梯度比较,确定切割位点。
与以前的发现(Zamore等人,2000)一致,所有靶RNA切割位点都位于与dsRNA相同的区域内。有义或反义靶标只被39bp dsRNA切割一次。每个切割位点都与dsRNA覆盖的区域的5’端相距10nt(图3B)。与39bp dsRNA有相同5’端的52bp dsRNA除了位于第一个位点下游23和24nt的两个较弱切割位点之外,在有义靶标上还产生相同的切割位点,距与dsRNA相同的区域的5’端10nt。反义靶标只切割一次,与各自dsRNA覆盖的区域的5’端相距10nt。图1所示的38-49bp dsRNA的切割位点作图显示,第一个和主要的切割位点总是位于dsRNA覆盖的区域的7-10nt下游(数据未显示)。这提示,靶RNA切割点取决于dsRNA末端,可能意味着加工为21-23mer从双链体末端开始。
较长的111bp dsRNA在有义和反义靶标上的切割位点比预期的更多,大多数似乎以20-23nt的间隔聚簇(图3A和3B)。对于较短的dsRNA,在有义靶标上的第一个切割位点与dsRNA覆盖的区域的5’端相距10nt,在反义靶标上的第一个切割位点与dsRNA覆盖的区域的5’端相距9nt。还不清楚是什么导致这种混乱的切割,但是一种可能性是,较长的dsRNA不仅可以从末端开始加工,而且也可在内部加工,或者存在我们还不清楚的一些dsRNA加工的特异性决定簇。以前也注意到21-23nt间隔的不规则性(Zamore等人,2000)。为了更好地了解dsRNA加工和靶RNA识别的分子基础,我们决定分析果蝇裂解液中39、52和111bp dsRNA加工产生的21-23nt片段的序列。
1.2.3 dsRNA通过RNase III样机制加工为21和22nt RNA
为了表征21-23nt RNA片段,我们检查了该RNA片段的5’和3’端。对凝胶纯化的21-23nt RNA进行高碘酸盐氧化,随后进行β-排除,表明存在末端2’和3’羟基。21-23mer对碱性磷酸酶处理也有反应性,表明存在5’端磷酸基团。5’磷酸和3’羟基端的存在提示,dsRNA能被类似于大肠杆菌RNase III的酶活性加工(综述见Dunn,1982;Nicholson,1999;Robertson,1990;Robertson,1982)。
利用T4 RNA连接酶将3’和5’接头寡核苷酸与纯化的21-23mer连接,进行21-23nt RNA片段的定向克隆。反转录连接产物,PCR扩增,连环化,克隆并测序。从39、52和111bp dsRNA的dsRNA加工反应中测序超过220个短RNA(图4A)。我们发现下列长度分布:1%18nt,5%19nt,12%20nt,45%21nt,28%22nt,6%23nt,2%24nt。对加工片段5’端核苷酸的序列分析表明,含有5’鸟苷的寡核苷酸被低估。这种情况最可能是由于加入了T4 RNA连接酶,该酶将5’磷酸化鸟苷识别为供体寡核苷酸;在3’端未见明显的序列偏差。来源于双链体有义或反义链3’端的许多~21nt片段含有在使用T7RNA聚合酶合成RNA过程中非模板添加核苷酸产生的3’核苷酸。有意思的是,也克隆了相当数量的内源果蝇~21nt RNA,其中一些来自LTR和非LTR反转录转座子(数据未显示)。这与RNAi在转座子沉默中的作用一致。
~21nt RNA存在于覆盖完整dsRNA序列的聚簇基团中(图4A)。显然,加工反应通过产生交错的3’末端切割dsRNA,这是RNase III切割的另一个特征。对于39bp dsRNA,在每条包含突出3’端的dsRNA组成链中发现两簇~21nt RNA,而在有义和反义靶标上只检测到一个切割位点(图3A和3B)。如果~21nt片段在介导mRNA降解的复合物中作为单链指导RNA存在,可以假定至少存在两个靶切割位点,但这不是事实。这提示,~21nt RNA可能以双链形式存在于内切核酸酶复合物中,但是只有一条链能用于靶RNA识别和切割。根据~21nt双链体与核酸酶复合物结合的方向可以简单地确定~21nt链之一在靶切割中的用途。该方向取决于原始dsRNA的加工方向。
与39bp dsRNA相比,52bp和111bp dsRNA的~21mer簇尚未确定。这些簇覆盖25-30nt的区域,最可能代表~21nt双链体的几个不同的亚群,因此指导在几个邻近位点的靶切割。这些切割区仍然主要有20-23nt的间隔。决定怎样规则的dsRNA才能被加工为~21nt片段的原则尚不清楚,但是以前发现,使用尿苷能改变切割位点的大约21-23nt的间隔(Zamore等人,2000)。大肠杆菌RNase III切割dsRNA的特异性似乎主要由抗决定簇控制,即,在相对于切割位点的特定位置处排除某些特异碱基对(Zhang和Nicholson,1997)。
为了检测加工的~21nt RNA片段中存在糖-、碱基-还是帽-修饰,我们在裂解液中温育放射性标记的505bp Pp-luc dsRNA 1小时,分离~21nt产物,用P1或T2核酸酶消化为单核苷酸。然后用2D薄层层析法分析核苷酸混合物(图4B)。P1或T2消化显示,4种天然核糖核苷酸都没有修饰。我们以前曾经分析了~21nt片段中的腺苷-肌苷转化(温育2小时后),检测到程度较小(<0.7%)的脱氨基作用(Zamore等人,2000);在裂解液中温育较短时间(1小时)使肌苷部分减至几乎无法检测的水平。可切割磷酸二酯键3’的RNase T2产生核苷3’-磷酸和核苷3’,5’-二磷酸,因而表明存在5’端一磷酸。检测到全部4种核苷3’,5’-二磷酸,提示核苷酸间键的切割没有或只有较低的序列特异性。总之,~21nt片段没有修饰,是由dsRNA产生的,因此在5’端存在5’-一磷酸和3’-羟基。
1.2.4合成21和22nt RNA介导靶RNA切割
对dsRNA加工产物的分析表明,~21nt片段是通过具有RNase III切割反应所有特征的一种反应产生的(Dunn,1982;Nicholson,1999;Robertson,1990;Robertson,1982)。RNase III在dsRNA的两条链上产生两个交错的切口,形成大约2nt的3’突出端。我们化学合成了21和22nt RNA,它们在序列上与某些克隆的~21nt片段相同,并且检测了它们介导靶RNA降解的能力(图5A和5B)。21和22nt RNA双链体在裂解液中以100nM浓度温育,浓度比52bp对照dsRNA高10倍。在这种条件下,易于检测到靶RNA切割。21和22nt双链体的浓度从100nM降到10nM仍能引起靶RNA切割。然而,双链体浓度从100nM提高到1000nM不能进一步提高靶切割,这可能是由于裂解液内存在一种限制蛋白因子。
与不介导RNAi的29或30bp dsRNA不同,具有2-4nt突出3’端的21和22nt dsRNA介导靶RNA的高效降解(双链体1、3、4、6,图5A和5B)。平端21或22nt dsRNA(双链体2、5、7,图5A和5B)降解靶标的能力降低,表明突出的3’端对于RNA-蛋白质核酸酶复合物的重建至关重要。~21nt双链体与蛋白质成分的高亲和力结合可能需要单链突出端。5’端磷酸尽管在dsRNA加工后存在,但不是介导靶RNA切割所需的,在合成的短RNA中缺乏。
合成21和22nt双链体指导有义及反义靶标在短双链体覆盖的区域内切割。这是一个重要结果,因为形成两对~21nt片段簇的39bpdsRNA(图2)只切割有义或反义靶标一次而不是两次。我们提出~21nt双链体的两条链只有一条能指导靶RNA切割,核酸酶复合物中~21nt双链体的方向取决于dsRNA加工的最初方向,从而解释了这一结果。然而,向体外系统输送已经加工好的~21nt双链体可以以对称RNA双链体的两个可能方向形成活性序列-特异性核酸酶复合物。这导致有义及反义靶标在与21nt RNA双链体相同的区域内切割。
靶切割位点位于与21或22nt指导序列互补的第一个核苷酸的11或12nt下游,即切割位点靠近21或22nt RNA所覆盖的区域的中心(图4A和4B)。替换22nt双链体有义链的两个核苷酸(比较图5A中的双链体1和3)只将反义靶标的切割位点替换了两个核苷酸。替换有义和反义链的两个核苷酸使切割位点位移两个核苷酸(比较双链体1和4)。我们预测能设计一对21或22nt RNA,它们几乎在任何特定位点均能切割靶RNA。
21和22nt RNA指导的靶RNA切割的特异性似乎是强烈的,因为未检测到异常的切割位点(图5B)。然而应当指出,21和22nt RNA双链体3’突出端存在的核苷酸在底物识别方面的作用可能比靠近切割位点的核苷酸要小。这是基于以下发现:活性双链体1或3的3’突出端中的3’核苷酸(图5A)不与靶标互补。现在利用21和22nt RNA能容易地进行RNAi特异性的详细分析。
根据具有突出3’端的21和22nt RNA介导RNA干涉的证据,我们提出将~21nt RNA命名为“短干涉RNA”或siRNA,将各自的RNA-蛋白质复合物命名为“小干涉核糖核蛋白颗粒”或siRNP。
1.2.5短dsRNA上的20nt 3’突出端抑制RNAi
我们表示,平端短dsRNA似乎从dsRNA的末端开始加工。在我们对RNAi中dsRNA长度依赖性的研究过程中,我们也分析了具有17-20nt突出3’端的dsRNA,我们吃惊地发现它们还不如平端dsRNA有效。对于可达100bp的dsRNA,长3’端的抑制作用特别明确,但是更长的dsRNA就没有这么明显。根据天然凝胶分析(数据未显示),这种作用是由于dsRNA形成不好。我们检验了长突出3’端的抑制作用是否能作为将dsRNA加工只定向于短RNA双链体两端之一的工具。
我们合成了52bp模型dsRNA的4种组合:平端、只在有义链上的3’延伸、只在反义链上的3’延伸和在两条链上的双3’延伸,并在裂解液中温育后对靶RNA切割位点作图(图6A和6B)。当双链体反义链的3’端延伸时,有义靶标的第一个和主要的切割位点丢失,反之亦然,当双链体有义链的3’端延伸时,反义靶标的强切割位点丢失。两条链的3’延伸使52bp dsRNA实际上失活。~20nt 3’延伸使dsRNA失活的一种解释是单链RNA-结合蛋白的结合,这能干扰dsRNA加工因子之一在该末端的结合。该结果也与我们的模型一致,在我们的模型中,在装配的siRNP中只有siRNA双链体的一条链能指导靶RNA切割。指导RNA切割的链的方向取决于dsRNA加工反应的方向。具有3’交错末端可能有利于加工复合物的装配。在有义链3’端的阻断只允许从反义链的相对3’端开始dsRNA加工。随后产生siRNP复合物,其中只有siRNA双链体的反义链能指导有义靶RNA切割。情况相反时同样如此。
至于较长的dsRNA(≥500bp,数据未显示),长3’延伸的抑制作用不明显,这提示我们,长dsRNA也可能含有内部dsRNA加工信号,或者可能由于多种切割因子的结合而协同加工。
1.2.6 dsRNA指导的mRNA切割的模型
新的生物化学数据更新了dsRNA如何靶向mRNA进行破坏的模型(图7)。双链RNA首先加工为主要长21和22nt的短RNA双链体,其具有类似于RNase III样反应的交错3’端(Dunn,1982;Nicholson,1999;Robertson,1982)。根据加工RNA片段的21-23nt长度,曾经推测RNase III样活性可能参与RNAi(Bass,2000)。在RNase III反应产物中发现,在siRNA的末端存在5’磷酸和3’羟基,这进一步支持了这一假说(Dunn,1982;Nicholson,1999)。细菌RNase III和真核同源物——酿酒酵母中的Rnt1p和粟酒酵母中的Pac1p在核糖体RNA和snRNA和snoRNA的加工中起作用(参见,例如Chanfreau等人,2000)。
关于来自植物、动物或人类的RNase III同源物的生物化学,人们知之甚少。主要通过数据库指导的序列分析或cDNA的克隆鉴定了两个RNase III酶家族。第一个RNase III家族的代表是1327个氨基酸长的黑尾果蝇蛋白drosha(Acc.AF116572)。C端由两个RNase III和一个dsRNA结合域组成,N端功能未知。在C.elegans(Acc.AF160248)和人类(Acc.AF189011)中也发现了密切同源物(Filippov等人,2000;Wu等人,2000)。drosha样人RNase III(Wu等人,2000)最近已经克隆并表征。该基因在人类组织和细胞系中普遍表达,该蛋白质位于细胞的核和核仁中。根据反义抑制研究推断的结果,提出了该蛋白质在rRNA加工中的作用。第二类的代表是编码1822个氨基酸长的蛋白质的C.elegans基因K12H4.8(Acc.S44849)。该蛋白质含有一个N端RNA解旋酶基序,之后是2个RNase III催化域和一个dsRNA结合基序,类似于drosha RNase III家族。在粟酒酵母(Acc.Q09884)、A.thaliana(Acc.FA187317)、黑尾果蝇(Acc.AE003740)和人类(Acc.AB028449)中具有密切同源物(Filippov等人,2000;Jacobsen等人,1999;Matsuda等人,2000)。K12H4.8 RNase III/解旋酶可能是参与RNAi的候选物。
在C.elegans中进行遗传筛选鉴定了rde-1和rde-4,它们是RNAi激活所必需的,对转座子移动或共抑制没有影响(Dernburg等人,2000;Grishok等人,2000;Ketting和Plasterk,2000;Tabara等人,1999)。这引出一种假说:这些基因对于dsRNA加工非常重要,但是不参与mRNA靶降解。这两种基因的功能仍然未知,rde-1基因产物是类似于兔蛋白elF2C的蛋白质家族的一员(Tabara等人,1999),rde-4的序列还未有描述。将来对这些蛋白质的生物化学表征将揭示其分子功能。
加工为siRNA双链体似乎从平端dsRNA或具有短(1-5nt)3’突出端的dsRNA的末端开始,以大约21-23nt的幅度加工。短dsRNA上的长(~20nt)3’交错末端抑制RNAi,这可能是通过与单链RNA结合蛋白相互作用。在短dsRNA侧翼的单链区抑制RNAi,以及短30bpdsRNA缺乏siRNA形成,可以解释mRNA中常见的结构区为什么不导致RNAi激活。
不希望被理论所束缚,我们假设dsRNA加工蛋白或其亚组在加工反应后仍与siRNA双链体结合。siRNA双链体相对于这些蛋白质的方向决定两条互补链的哪一条在指导靶RNA降解中起作用。化学合成的siRNA双链体指导有义及反义靶RNA的切割,因为它们能以两种可能的方向之一与蛋白质成分结合。
合成21和22nt siRNA双链体能用于有效的mRNA降解,这一重要发现为功能基因组学以及生物医学研究中基因表达的序列特异性调节提供了新的工具。在由于PKR反应激活而不能使用长dsRNA的哺乳动物系统中,siRNA可能有效(Clemens,1997)。因此,siRNA双链体是反义或核酶治疗剂的一种新的备选剂。
实施例2
人组织培养物中的RNA干涉
2.1方法
2.1.1RNA制备
用Expedite RNA亚磷酰胺和胸苷亚磷酰胺(Proligo,德国)化学合成21nt RNA。合成的寡核苷酸去保护并凝胶纯化(实施例1),之后经Sep-Pak C18柱(Waters,Milford,MA,USA)纯化(Tuschl,1993)。针对GL2(Acc.X65324)和GL3萤光素酶(Acc.U47296)的siRNA序列对应于相对于起始密码子第一个核苷酸的编码区152-173,针对RL(Acc.AF025846)的siRNA对应于起始密码子之后的区119-129。较长的RNA用T7 RNA聚合酶由PCR产物转录,然后用凝胶和Sep-Pak纯化。49和484bp GL2或GL3 dsRNA分别对应于相对于翻译起点的位点113-161和113-596。50和501bp RL dsRNA分别对应于位点118-167和118-618。针对人源化GFP(hG)的dsRNA合成的PCR模板从pAD3(Kehlenbach,1998)中扩增,因此50和501bp hG dsRNA分别对应于相对起始密码子的位点118-167和118-618。
为使siRNA退火,20μM单链在退火缓冲液(100mM乙酸钾,30mM HEPES-KOH pH7.4,2mM乙酸镁)中90℃温育1分钟,然后37℃ 1小时。对于50和500bp dsRNA,37℃温育步骤延长过夜,退火反应分别以8.4μM和0.84μM的链浓度进行。
2.1.2细胞培养
S2细胞在25℃下在补充了10%FBS、100单位/ml青霉素和100μg/ml链霉素的Schneider’s果蝇培养基(Life Technologies)中繁殖。293、NIH/3T3、HeLa S3、COS-7细胞在37℃下在补充了10%FBS、100单位/ml青霉素和100μg/ml链霉素的Dulbecco’s改良Eagle’s培养基中培养。细胞定期传代,以保持指数生长。转染前24小时,在大约80%平铺时,哺乳动物细胞用胰蛋白酶消化,用不含抗生素的新鲜培养基1∶5稀释(1-3×105细胞/ml),转移到24孔板中(500μl/孔)。S2细胞在分裂前不用胰蛋白酶消化。如厂商对于粘附细胞系所述,用Lipofectamine 2000试剂(Life Technologies)进行转染。每孔加入配制于脂质体中的1.0μg pGL2-对照(Promega)或pGL3-对照(Promega)、0.1μg pRL-TK(Promega)和0.28μg siRNA双链体或dsRNA;终体积为每孔600μl。细胞在转染20小时后温育,此后似乎健康。然后用双萤光素酶测定(Promega)监测萤光素酶表达。在1.1μg hGFP编码pAD3和0.28μg invGL2 inGL2 siRNA共转染后,通过对哺乳动物细胞系的荧光显微镜检,测定转染率,为70-90%。报道质粒在XL-1 Blue(Stratagene)中扩增,用Qiagen EndoFree Maxi质粒试剂盒纯化。
2.2结果与讨论
为了检验siRNA是否也能在组织培养中介导RNAi,我们合成了具有对称2nt 3’突出端的21nt siRNA双链体,它们针对编码海肾(Renilla reniformis)和萤火虫(Photinus pyralis,GL2和GL3)萤光素酶两种序列变体的报道基因(图8a,b)。siRNA双链体与报道质粒组合pGL2/pRL或pGL3/pRL利用阳离子脂质体共转染黑尾果蝇Schneider S2细胞或哺乳动物细胞。在转染20小时后测定萤光素酶活性。在检测的所有细胞系中,我们在同源siRNA双链体存在下观察到报道基因表达特异性减少(图9a-j)。显然,绝对萤光素酶表达水平不受非同源siRNA影响,表明21nt RNA双链体没有有害的副作用(对于HeLa细胞,例如图10a-d)。在黑尾果蝇Schneider S2细胞中(图9a,b),萤光素酶完全特异性抑制。在报道基因表达强50-100倍的哺乳动物细胞中,特异性抑制不完全(图9c-j)。同源siRNA致使GL2表达减少3-12倍,GL3表达减少9-25倍,RL表达减少1-3倍。对于293细胞,RLsiRNA对RL萤光素酶的靶向无效,但是GL2和GL3靶特异性反应(图9i,j)。293细胞中没有RL表达减少可能是由于其表达比检测的其它任何哺乳动物细胞系高5-20倍,和/或由于RNA二级结构或结合蛋白质,靶序列的可接近性受限。然而,同源siRNA双链体对GL2和GL3萤光素酶的特异靶向表明RNAi在293细胞中也起作用。
除了uGL2之外,所有siRNA双链体的2nt 3’突出端都由(2’-脱氧)胸苷组成。3’突出端中尿苷置换为胸苷在黑尾果蝇体外系统中良好耐受,突出端序列对靶标识别而言并不重要。选择胸苷突出端,因为推断它能提高组织培养基中和转染细胞内siRNA的核酸酶抗性。实际上,在检测的所有细胞系中,胸苷修饰的GL2 siRNA略强于未修饰的uGL2 siRNA(图9a,c,e,g,i)。可以想象,进一步修饰3’突出端核苷酸可能对siRNA双链体的输送和稳定性有其它好处。
在共转染实验中,对于组织培养基的终体积,使用25nM siRNA双链体(图9,10)。siRNA浓度提高为100nM不能提高特异性沉默作用,但是由于质粒DNA与siRNA之间脂质体包封的竞争,开始影响转染率(数据未显示)。siRNA浓度降至1.5nM不会降低特异性沉默作用(数据未显示),即使siRNA只比DNA质粒浓缩2-20倍。这表明,siRNA是介导基因沉默的非常有效的试剂,在浓度比常规反义或核酶基因靶向实验使用的浓度低几个数量级时,siRNA是有效的。
为了监测较长dsRNA对哺乳动物细胞的作用,制备了与报道基因同源的50和500bp dsRNA。用来自人源化GFP(hG)的dsRNA作为非特异性对照(Kehlenbach,1998)。当dsRNA与siRNA双链体含量相同地(不浓缩)共转染时,报道基因的表达强烈且非特异性地降低。以HeLa细胞为代表性实例说明了这一结果(图10a-d)。50bp dsRNA共转染使绝对萤光素酶活性非特异地降低10-20倍,500bp dsRNA共转染使其降低20-200倍。对于COS-7和NIH/3T3细胞也观察到类似的非特异性作用。对于293细胞,只有用500bp dsRNA才观察到10-20倍非特异性降低。>30bp dsRNA使报道基因表达非特异性降低被认为是干扰素反应的一部分。
奇怪的是,尽管报道基因表达强烈地非特异性降低,但我们可重复地检测到其它序列特异的、dsRNA介导的沉默。然而,只有在相对报道基因活性用hG dsRNA对照标准化时,特异性沉默作用才明显(图10e,f)。在检测的其它3种哺乳动物细胞系中也观察到同源dsRNA引起的2-10倍的特异性降低(数据未显示)。dsRNA(356-1662bp)的特异性沉默作用以前在CHO-K1细胞中有报道,但是检测2-4倍特异性降低所需的dsRNA的量比我们的实验大约高20倍(Ui-Tei,2000)。CHO-K1细胞的干扰素反应似乎也有缺陷。在另一篇报告中,利用萤光素酶/lacZ报道分子组合和829bp特异性lacZ或717bp非特异性GFP dsRNA检测293、NIH/3T3和BHK-21细胞的RNAi(Caplen,2000)。在这种情况下无法检测RNAi可能是由于萤光素酶/lacZ报道分子测定法的低敏感性和靶标与对照dsRNA的长度差异。我们的结果综合起来表明,RNAi在哺乳动物细胞中有活性,但是,如果干扰素系统被>30bp的dsRNA激活,则沉默作用难以检测。
总之,我们第一次在哺乳动物细胞中证明了siRNA介导的基因沉默。短siRNA的应用在人类组织培养的基因功能灭活和基因特异治疗剂的研制中具有广阔前景。
实施例3
RNA干涉对基因表达的特异性抑制
3.1材料与方法
3.1.1RNA制备和RNAi测定
化学RNA合成、退火和基于萤光素酶的RNAi测定如实施例1或2或以前的公开文献所述进行(Tuschl等人,1999;Zamore等人,2000)。所有siRNA双链体都针对萤火虫萤光素酶,萤光素酶mRNA序列来源于如(Tuschl等人,1999)所述的pGEM-luc(GenBank acc.X65316)。在加入mRNA之前,siRNA双链体在黑尾果蝇RNAi/翻译反应液中温育15分钟。基于翻译的RNAi测定至少一式三份进行。
为了对有义靶RNA切割作图,产生177-nt转录物,其对应于在相对于起始密码子的位点113-273之间的萤火虫萤光素酶序列,随后为SP6启动子序列的17-nt互补序列。为了对反义靶RNA切割作图,由模板产生166-nt转录物,它是利用5’引物TAATACGACTCACTATAGAGCCCATATCGTTTCATA(下划线标出T7启动子)和3’引物AGAGGATGGAACCGCTGG由质粒序列PCR扩增的。靶序列对应于在相对于起始密码子的位点50-215之间的萤火虫萤光素酶序列的互补序列。鸟苷酰基转移酶标记如前所述进行(Zamore等人,2000)。为了对靶RNA切割作图,在标准条件下(Zamore等人,2000),100nM siRNA双链体与5-10nM靶RNA在黑尾果蝇胚胎裂解液中25℃温育2小时。加入8倍体积的蛋白酶K缓冲液(200mMTris-Hcl pH7.5,25mM EDTA,300mM NaCl,2%w/v十二烷基硫酸钠)终止反应。加入蛋白酶K(E.M.Merck,溶于水)至终浓度为0.6mg/ml。反应液然后在65℃下温育15分钟,用苯酚/氯仿/异戊醇(25∶24∶1)抽提,用3倍体积的乙醇沉淀。样品置于6%测序凝胶上。长度标准通过帽标记的有义或反义靶RNA的部分RNase T1消化和部分碱水解产生。
3.2结果
3.2.1 21nt siRNA双链体中3’突出端的变异
如上所述,位于siRNA双链体3’末端的2个或3个未配对核苷酸在靶RNA降解上比相应的平端双链体更有效。为了更全面地分析末端核苷酸的功能,我们合成了5种21nt有义siRNA,每一个均用相对于靶RNA的一个核苷酸展示,而且合成了8种21nt反义siRNA,每一个均用相对于靶标的一个核苷酸展示(图11A)。通过组合有义和反义siRNA,产生8个系列的siRNA双链体,它们具有合成的突出末端,覆盖7nt 3’突出端到4nt 5’突出端的范围。用双萤光素酶测定系统测定siRNA双链体的干涉(Tuschl等人,1999;Zamore等人,2000)。siRNA双链体针对萤火虫萤光素酶mRNA,海肾萤光素酶mRNA作为内部控制。在siRNA双链体存在下测定靶标与对照萤光素酶活性的发光比,用不含dsRNA时的发光比标准化。为了比较,在图11B中显示长dsRNA(39-504bp)的干涉比。长dsRNA在5nM浓度下(图11A),siRNA双链体在100nM浓度下(图11C-J)测定干涉比。选择100nM的siRNA浓度,因为5nM 504bp dsRNA的完全加工将产生120nM总siRNA双链体。
21nt siRNA双链体介导RNAi的能力依赖于突出核苷酸或形成的碱基对的数量。含有4-6个3’突出核苷酸的双链体不能介导RNAi(图11C-F),含有2个或2个以上5’突出核苷酸的双链体同样不能(图11G-J)。具有2nt 3’突出端的双链体在介导RNA干涉上最有效,但是沉默率也是序列依赖性的,具有2nt 3’突出端的不同siRNA双链体观察到高达12倍的差异(比较图11D-H)。具有平端、1nt 5’突出端或1-3nt 3’突出端的双链体有时有功能。具有7nt 3’突出端的siRNA双链体所观察到的较小的沉默作用(图11C)可能是由于长3’突出端的反义作用,而不是由于RNAi。长dsRNA(图11B)与最有效的21nt siRNA双链体(图11E,G,H)之间RNAi效率的比较表明,100nM浓度的单siRNA双链体能与5nM 504bp dsRNA一样有效。
3.2.2与恒定21-nt反义siRNA配对的有义siRNA的长度变化
为了研究siRNA长度对RNAi的影响,我们制备了3个系列的siRNA双链体,使3种21nt反义链与8种18-25nt有义链结合。在每个siRNA双链体系列中,反义siRNA的3’突出端固定为1、2或3nt,而有义siRNA的3’端不同(图12A)。我们发现,与有义siRNA的长度无关,具有反义siRNA的2-nt 3’突出端的双链体(图12C)比具有1-nt或3-nt 3’突出端的双链体更有活性(图12B,D)。在具有反义siRNA的1-nt 3’突出端的第一个系列中,含有21-nt和22-nt有义siRNA的双链体(它们分别携带有义siRNA的1-nt和2-nt 3’突出端)最具活性。含有19-25nt有义siRNA的双链体也能介导RNA,但程度较低。类似地,在具有反义siRNA的2-nt突出端的第二个系列中,具有2-nt 3’突出端的21-nt siRNA双链体最具活性,其他任何与18-25nt有义siRNA的结合都有显著的活性。在具有3-nt反义siRNA 3’突出端的最后一个系列中,只有含20-nt有义siRNA和2-nt有义3’突出端的双链体能减少靶RNA表达。这些结果综合起来表明,siRNA的长度以及3’突出端的长度非常重要,具有2-nt 3’突出端的21-nt siRNA双链体最适于RNAi。
3.2.3具有恒定2-nt 3’突出端的siRNA双链体的长度变化
然后我们检查了通过保留对称2-nt 3’突出端同时改变两条siRNA链长度的影响(图13A)。制备两个系列的siRNA双链体,包括图11H的21-nt siRNA双链体作为参照。通过在有义siRNA的3’端(图13B)或在反义siRNA的3’端(图13C)延长碱基配对的片段,双链体的长度在20-25bp间不等。20-23bp双链体引起靶萤光素酶活性的特异性抑制,但是21-nt siRNA双链体比其它任何双链体有效性至少高8倍。24-和25-nt siRNA双链体不引起任何可检测的干涉。由于双链体两端的变化产生类似的作用,序列特异的影响较小。
3.2.4 2’-脱氧和2’-O-甲基修饰的siRNA双链体
为了评估siRNA核糖残基对于RNAi的重要性,检测了含有21-ntsiRNA和2-nt 3’突出端以及2’-脱氧或2’-O-甲基修饰的链的双链体(图14)。2-nt 3’突出端置换为2’-脱氧核苷酸没有影响,甚至置换配对区内与突出端相邻的另外两个核糖核苷酸产生具有明显活性的siRNA。因此,siRNA双链体的42nt中的8nt置换为DNA残基,活性没有损失。然而,一条或两条siRNA链完全置换为2’-脱氧残基消除了RNAi,置换为2’-O-甲基残基同样如此。
3.2.5靶RNA切割位点的确定
以前确定了22-nt siRNA双链体和21-nt/22-nt双链体的靶RNA切割位点。发现靶RNA切割位点位于siRNA双链体覆盖的区域的中心,在与21-或22-nt siRNA指导序列互补的第一个核苷酸的11或12nt下游。具有2-nt 3’突出端的5种不同21-nt siRNA双链体(图15A)与5’帽标记的有义或反义靶RNA在黑尾果蝇裂解液中温育(Tuschl等人,1999;Zamore等人,2000)。5’切割产物在测序凝胶上分析(图15B)。切割的有义靶RNA的量与通过基于翻译的实验测定的siRNA双链体的效率有关,siRNA双链体1、2、4(图15B和11H,G,E)切割靶RNA比双链体3和5更快(图15B和11F,D)。显然,5’切割产物和输入靶RNA的总放射性随着时间的增长不是恒定的,而且5’切割产物不积累。推测起来,由于缺乏5’-帽子的任一条poly(A)尾,切割产物一从siRNA-内切核酸酶复合物上释放,即快速降解。
有义和反义靶RNA的切割位点都位于siRNA双链体覆盖的区域的中间。根据双链体沿靶序列的1-nt置换来看,5种不同双链体产生的每种靶标的切割位点有1-nt不同。靶标精确地在靶位点11nt下游切割,靶位点与序列互补的指导siRNA的3’-核苷酸互补(图15A,B)。
为了确定是指导siRNA的5’端还是3’端控制靶RNA切割,我们设计了实验策略,在图16A和B中概述。一种21-nt反义siRNA,在该研究中保持不变,与5’或3’端修饰的有义siRNA配对。有义和反义靶RNA的切割位点如上所述确定。有义siRNA的3’端的改变(监测1-nt5’突出端到6-nt 3’突出端)不影响有义或反义靶RNA的切割位点(图16C)。有义siRNA的5’端的改变不影响有义靶RNA的切割(图16D,上图),预计是因为反义siRNA未改变。然而,反义靶RNA切割受到影响,并且强烈依赖于有义siRNA的5’端(图16D,下图)。当有义siRNA大小为20或21nt时,只切割反义靶标,切割位点时相差1-nt,表明靶标识别的siRNA的5’端控制靶RNA的切割。当从与指导siRNA的5’核苷酸配对的靶核苷酸开始以下游方向计数时,该位点位于核苷酸10与11之间(参见图15A)。
3.2.6序列效应和3’突出端的2’-脱氧置换
对于siRNA的功能,2-nt 3’突出端是优选的。我们希望了解突出核苷酸的序列对于靶标识别是否有作用,或者这是否只是内切核酸酶复合物(RISC或siRNP)重建所需的一个特征。我们合成了有义和反义siRNA,其具有AA、CC、GG、UU和UG 3’突出端,包括2’-脱氧修饰TdG和TT。野生型siRNA在有义3’突出端中含有AA,在反义3’突出端中含有UG(AA/UG)。所有siRNA双链体在干涉测定中都起作用,使靶表达至少降低5倍(图17)。最有效的siRNA双链体是序列型NN/UG、NN/UU、NN、TdG和NN/TT(N,任何核苷酸),使靶表达降低10倍以上。含有AA、CC或GG的反义siRNA 3’突出端的siRNA双链体比野生型序列UG或突变UU活性低2-4倍。这种RNAi效率的降低可能是由于倒数第二个3’核苷酸对序列特异性靶标识别的作用,因为3’端核苷酸由G变为U没有影响。
有义siRNA 3’突出端序列的改变没有显示任何依赖序列的作用,这是预料到的,因为有义siRNA对于有义靶mRNA的识别不一定有作用。
3.2.7靶标识别的序列特异性
为了检验靶标识别的序列特异性,我们向siRNA双链体的配对片段内引入序列改变,并测定沉默率。通过颠倒3-或4-nt长的短片段或点突变,引入序列改变(图18)。为了避免干扰碱基配对的siRNA双链体结构,一条siRNA链的序列改变在互补siRNA链中补偿。为了降低合成成本,所有2-nt 3’突出端序列都是TT(T,2’-脱氧胸苷)。TT/TT参照siRNA双链体在RNAi方面与野生型siRNA双链体AA/UG相当(图17)。用基于翻译的发光测定法定量介导报道mRNA破坏的能力。显示含有反向序列片段的siRNA双链体靶向萤火虫萤光素酶报道分子的能力显著降低(图18)。位于反义siRNA 3’端与中心之间的序列改变完全消除了靶RNA识别,但是靠近反义siRNA 5’端的突变显示程度较低的沉默。与预测的靶RNA切割位点正对的A/U碱基对颠换,或者远离预测位点的一个核苷酸的颠换,阻止靶RNA的切割,因此表明,siRNA双链体中心的单突变区别错配的靶标。
3.3讨论
siRNA是不仅在昆虫细胞中,而且在哺乳动物细胞中,用于灭活基因表达的有价值的试剂,具有广阔的治疗应用前景。我们系统分析了在黑尾果蝇胚胎裂解液中促进高效靶RNA降解所需的siRNA双链体的结构决定簇,从而提出了最有效的siRNA双链体的设计原则。完美的siRNA双链体能够沉默基因表达,假定使用含量相当的总RNA,其效率相当于500bp dsRNA。
3.4siRNA使用指南
高效沉默的siRNA双链体优选地由21-nt反义siRNA组成,应当选择它们形成具有2-nt 3’突出端的19bp双螺旋。2-nt 3’突出核糖核苷酸的2’-脱氧置换不影响RNAi,但是有助于降低RNA合成的成本,并且可提高siRNA双链体的RNAse抗性。然而,更广泛的2’-脱氧或2’-O-甲基修饰降低了siRNA介导RNAi的能力,这可能是通过干扰SiRNAP装配的蛋白质结合。
靶标识别是一个高度序列特异性的过程,由与靶标互补的siRNA介导。指导siRNA的3’-核苷酸对于靶标识别的特异性没有作用,而3’突出端的倒数第二个核苷酸影响靶RNA切割,错配使RNAi降低2-4倍。指导siRNA的5’端似乎也比3’端更能容许错配的靶RNA识别。位于靶RNA切割位点对面的siRNA中心的核苷酸是重要的特异性决定簇,甚至单核苷酸改变也能使RNAi降至不可检测的水平。这提示,在基因靶向实验中,siRNA双链体能区别突变的或多态性等位基因,这可能成为未来治疗发展的重要特征。
当有义和反义siRNA与内切核酸酶复合物或其定型复合物的蛋白质成分结合时,认为两者起不同的作用;该复合物中siRNA双链体的相对方向决定了哪条链能用于靶标识别。合成siRNA双链体在双螺旋结构上二重对称,但序列不对称。siRNA双链体在黑尾果蝇裂解液中与RNAi蛋白的结合将形成两种不对称复合物。在这种假定的复合物中,有义和反义siRNA的手性环境不同,因此功能不同。这种预测显然不适用于回文siRNA序列,或者能结合为同型二聚体的RNAi蛋白。为了使可影响有义与反义靶向siRNP之比的序列效应达到最小,我们建议使用含有相同3’突出序列的siRNA序列。我们推荐将有义siRNA突出端的序列调节为反义3’突出端的序列,因为有义siRNA在一般的敲除实验中没有靶标。有义和反义切割的siRNP重建的不对称性可能(部分地)导致RNAi效率的改变,用该研究使用的具有2-nt 3’突出端的多种21-nt siRNA双链体观察到这种改变(图14)。此外,靶位点处的核苷酸序列和/或靶RNA结构的可接近性可能导致这些siRNA双链体的效率的改变。
参考文献
Bass,B.L.(2000).双链RNA作为基因沉默的模板。Cell 101,235-238.
Bosher,J.M.和Labouesse,M.(2000).RNA干涉:遗传魔杖和遗传看家狗.Nat.Cell Biol.2,E31-36.
Caplen,N.J.,Fleenor,J.,Fire,A.和Morgan,R.A.(2000).培养的果蝇细胞中dsRNA-介导的基因沉默:用于分析RNA干涉的一种组织培养模型。Gene 252,95-105.
Catalanotto,C.,Azzalin,G.,Macino,G.和Cogoni,C.(2000).蠕虫和真菌中的基因沉默。Nature 404,245.
Chanfreau,G.,Buckle,M.和Jacquier,A.(2000).酿酒酵母RNaseIII对一类保守的RNA四环的识别。Proc.Natl.Acad.Sci.USA 97,3142-3147.
Clemens,M.J.(1997).PKR-由双链RNA调节的一种蛋白激酶。Int.J.Biochem.Cell Biol.29,945-949.
Cogoni,C.和Macino,G.(1999).植物和真菌中依赖同源性的基因沉默:同一主题的大量变异。Curr.Opin.Microbiol.2,657-662.
Dalmay,T.,Hamilton,A.,Rudd,S.,Angell,S.和Baulcombe,D.C.(2000).转基因介导的而不是病毒介导的转录后基因沉默需要Arabidopsis的一种RNA-依赖性RNA聚合酶基因。Cell 101,543-553.
Dernburg,A.F.,Zalevsky,J.,Colaiacovo,M.P.和Villeneuve,A.M.(2000).C.elegans种系中转基因介导的共抑制。Genes & Dev.14,1578-1583.
Dunn J.J.(1982)。核糖核酸酶III。《酶》,第15卷,部分B,P.D.Boyer编著(New York:Academic Press),pp.485-499.
Filippov,V.,Solovyev,V.,Filippova,M.和Gill,S.S.(2000).真核生物中的一种新型RNase III家族蛋白。Gene 245,213-221.
Fire,A.(1999).RNA-触发的基因沉默。Trends Genet.15,358-363.
Fire,A.,Xu,S.,Montgomery,M.K.,Kostas,S.A.,Driver,S.E.和Mello,C.C.(1998).Caenorhabditis elegans中双链RNA引起的强烈特异性遗传干涉。Nature 391,806-811.
Grishok,A.,Tabara,H.和Mello,C.C.(2000).C.elegans中RNAi遗传的基因要求。Science 287,2494-2497.
Hamilton,A.J.和Baulcombe,D.C.(1999).植物转录后基因沉默中的一个小反义RNA种。Science 286,950-952.
Hammond,S.M.,Bernstein,E.,Beach,D.和Hannon,G.J.(2000).在果蝇细胞中,一种RNA指导的核酸酶介导转录后基因沉默。Nature404,293-296.
Jacobsen,S.E.,Running,M.P.和M.,M.E.(1999).Arabidopsis中RNA解旋酶/RNase III基因的破坏导致花分生组织中无控制的细胞分裂。Development 126,5231-5243.
Jensen,S.,Gassama,M.P.和Heidmann,T.(1999).依赖同源性的基因沉默对转座因子的抑制(Taming)。Nat.Genet.21,209-212.
Kehlenbach,R.H.,Dickmanns,A.& Gerace,L.(1998).包括Ran和CRM1在内的核胞质穿梭因子介导NFAT的体外核输出。J.CellBiol.141,863-874.
Kennerdell,J.R.和Carthew,R.W.(1998).dsRNA-介导的遗传干涉证明无翅途径中frizzled和frizzled 2作用的用途。Cell 95,1017-1026.
Ketting,R.F.,Haverkamp,T.H.,van Luenen,H.G.和Plasterk,R.H.(1999).转座子沉默和RNA干涉需要的C.elegans的Mut-7是werner综合征解旋酶和RNaseD的同源物。Cell 99,133-141.
Ketting,R.F.和Plasterk,R.H.(2000).C.elegans中共抑制与RNA干涉之间的遗传联系。Nature 404,296-298.
Lucy,A.P.,Guo,H.S.,Li,W.X.和Ding,S.W.(2000).位于核中的植物病毒蛋白对转录后基因沉默的抑制。EMBO J.19,1672-1680.
Matsuda,S.,Ichigotani,Y.,Okuda,T.,Irimura,T.,Nakatsugawa,S.和Hamaguchi,M.(2000).编码推断的RNA解旋酶的一种新人类基因(HERNA)的分子克隆和表征。Biochim.Biophys.Acta 31,1-2.
Milligan,J.F.和Uhlenbeck,O.C.(1989).利用T7 RNA聚合酶合成小RNAs。Methods Enzymol.180,51-62.
Mourrain,P.,Beclin,C.,Elmayan,T.,Feuerbach,F.,Godon,C.,Morel,J.B.,Jouette,D.,Lacombe,A.M.,Nikic,S.,Picault,N.,Remoue,K.,Sanial,M.,Vo,T.A.和Vaucheret,H.(2000).转录后基因沉默和天然病毒抗性需要Arabidopsis SGS2和SGS3基因。Cell 101,533-542.
Ngo,H.,Tschudi,C.,Gull,K.和Ullu,E.(1998).在布氏锥虫(Trypanosoma brucei)中双链RNA诱导mRNA降解。Proc.Natl.Acad.Sci.USA 95,14687-14692.
Nicholson,A.W.(1999).细胞核糖核酸酶的功能、机制和调节。FEMS Microbiol.Rev.23,371-390.
Oelgeschlager,M.,Larrain,J.,Geissert,D.和De Robertis,E.M.(2000).进化上保守的BMP-结合蛋白缠绕的原肠胚形成促进BMP信号发生。Nature 405,757-763.
Pan,T.和Uhlenbeck,O.C.(1992).经历Pb2+自切割的RNAs的体外选择。Biochemistry 31,3887-3895.
Pelissier,T.和Wassenegger,M.(2000).一种30bp的DNA靶标对于RNA-指导的甲基化是足够的。RNA 6,55-65.
Plasterk,R.H.和Ketting,R.F.(2000).基因沉默。Curr.Opin.Genet.Dev.10,562-567.
Ratcliff,F.G.,MacFarlane,S.A.和Baulcombe,D.C.(1999).不含DNA的基因沉默。病毒间RNA介导的交叉保护。Plant Ceu 11.1207-1216.
Robertson,H.D.(1990).大肠杆菌核糖核酸酶III。MethodsEnzymol.181,189-202.
Robertson,H.D.(1982).大肠杆菌核糖核酸酶III切割位点。Cell30,669-672.
Romaniuk,E.,McLaughlin,L.W.,Neilson,T.和Romaniuk,P.J.(1982).受体寡核糖核苷酸序列对T4 RNA连接酶反应的影响。Eur JBiochem 125,639-643.
Sharp,P.A.(1999).RNAi和双链RNA.Genes & Dev.13,139-141.
Sijen,T.和Kooter,J.M.(2000).转录后基因沉默:RNAs是攻击不是防御?Bioessays 22,520-531.
Smardon,A.,Spoerke,J.,Stacey,S.,Klein,M.,Mackin,N.和Maine,E.(2000).在C.elegans中,EGO-1与RNA指导的聚合酶和种系发育中的功能和RNA干涉有关。Curr.Biol.10,169-178.
Svoboda,P.,Stein,P.,Hayashi,H.和Schultz,R.M.(2000).RNA干涉所致小鼠卵母细胞中静止母本mRNAs的选择性减少。Development 127,4147-4156.
Tabara,H.,Sarkissian,M.,Kelly,W.G.,Fleenor,J.,Grishok,A.,Timmons,L.,Fire,A.和Mello,C.C.(1999).C.elegans中的rde-1基因、RNA干涉和转座子沉默。Cell 99,123-132.
Tuschl,T.,Ng,M.M.,Pieken,W.,Benseler,F.和Eckstein,F.(1993).核心鸟苷的外环碱基功能基团对于锤头状核酶活性的重要性。Biochemistry 32,11658-11668.
Tuschl,T.,Sharp,P.A.和Bartel,D.P.(1998).从部分随机化的U2和U6 snRNA文库中体外选择新型核酶。EMBO J.17,2637-2650.
Tuschl,T.,Zamore,P.D.,Lehmann,R.,Bartel,D.P.和Sharp,P.A.(1999).体外双链RNA引起的定向mRNA降解。Genes & Dev.13,3191-3197.
Ui-Tei,K.,Zenno,S.,Miyata,Y.和Saigo,K.(2000).利用萤火虫萤光素酶基因作为靶标对果蝇和中国仓鼠培养细胞中RNA干涉的敏感测定。FEBS Letters 479,79-82.
Verma,S.和Eckstein,F.(1999).修饰的寡核苷酸:使用者的合成和策略。Annu.Rev.Biochem.67,99-134.
Voinnet,O.,Lederer,C.和Baulcombe,D.C.(2000).在烟草Nicotiana benthamiana中,一种病毒运动蛋白阻止基因沉默信号的扩散。Cell 103,157-167.
Wassenegger,M.(2000).RNA-指导的DNA甲基化 Plant Mol.Biol.43,203-220.
Wianny,F.和Zernicka-Goetz,M.(2000).早期小鼠发育过程中双链RNA对基因功能的特异性干涉。Nat.Cell Biol. 2,70-75.
Wu,H.,Xu,H.,Miraglia,L.J.和Crooke,S.T.(2000).人RNaseIII是一种参与前核糖体RNA加工的160kDa蛋白质。J.Biol.Chem.17,17.
Yang,D.,Lu,H.和Erickson,J.W.(2000)在果蝇胚胎中,在RNAi期间,加工的小dsRNAs可以介导序列特异的mRNA降解的证据。Curr.Biol.,10,1191-1200.
Zamore,P.D.,Tuschl,T.,Sharp,P.A.和Bartel,D.P.(2000).RNAi:双链RNA指导mRNA以21-23个核苷酸的间隔依赖ATP地切割。Cell 101,25-33.
Zhang,K.和Nicholson,A.W.(1997).双螺旋序列抗决定簇对核糖核酸酶III加工的调节。Proc.Natl.Acad.Sci.USA 94,13437-13441.

Claims (47)

1.分离的双链RNA分子,其中每条RNA链的长度为19-25个核苷酸,该RNA分子能够靶特异性核酸修饰。
2.权利要求1的RNA分子,其中至少一条链具有1-5个核苷酸的3’-突出端。
3.权利要求1或2的RNA分子,它能靶特异性RNA干涉和/或DNA甲基化。
4.权利要求1-3中任一项的RNA分子,其中每条链的长度为19-23个,特别是20-22个核苷酸。
5.权利要求2-4中任一项的RNA分子,其中3’-突出端为1-3个核苷酸。
6.权利要求2-5中任一项的RNA分子,其中为避免降解稳定3’-突出端。
7.权利要求1-6中任一项的RNA分子,它含有至少一个修饰的核苷酸类似物。
8.权利要求7的RNA分子,其中修饰的核苷酸类似物选自糖修饰或骨架修饰的核糖核苷酸。
9.根据权利要求7或8的RNA分子,其中该核苷酸类似物是糖修饰的核糖核苷酸,2’-OH基被置换为选自H、OR、R、卤素、SH、SR1、NH2、NHR、NR2或CN的基团,其中R是C1-C6烷基、烯基或炔基,卤素是F、Cl、Br或I。
10.权利要求7或8的RNA分子,其中该核苷酸类似物是含有硫代磷酸酯基的骨架修饰的核糖核苷酸。
11.权利要求1-10中任一项的RNA分子,其序列与预定的mRNA靶分子有至少50%的同一性。
12.权利要求11的RNA分子,其中同一性至少为70%。
13.一种制备权利要求1-12中任一项的双链RNA分子的方法,包括下列步骤:
(a)合成两条RNA链,每条长度为19-25个核苷酸,其中该RNA链能够形成双链RNA分子,
(b)在一定条件下结合合成的RNA链,其中形成一种双链RNA分子,它能靶特异性核酸修饰。
14.权利要求13的方法,其中化学合成RNA链。
15.权利要求13的方法,其中酶促合成RNA链。
16.一种介导细胞或生物中靶特异性核酸修饰的方法,包括下列步骤:
(a)在可发生靶特异性核酸修饰的条件下使细胞或生物接触权利要求1-12中任一项的双链RNA分子,和
(b)将双链RNA完成的靶特异性核酸修饰导向含有基本对应于该双链RNA的序列部分的靶核酸。
17.权利要求16的方法,其中核酸修饰是RNA干涉和/或DNA甲基化。
18.权利要求16和17的方法,其中所述接触包括向可发生靶特异性核酸修饰的靶细胞中导入所述双链RNA分子。
19.权利要求18的方法,其中这种导入包括载体介导的输送或注射。
20.权利要求16-19中任一项的方法的用途,用于确定细胞或生物中一种基因的功能。
21.权利要求16-19中任一项的方法的用途,用于调节细胞或生物中一种基因的功能。
22.权利要求20或21的用途,其中该基因与一种病理状态有关。
23.权利要求22的用途,其中该基因是一种病原体相关基因。
24.权利要求23的用途,其中该基因是一种病毒基因。
25.权利要求22的用途,其中该基因是一种肿瘤相关基因。
26.权利要求22的用途,其中该基因是一种自身免疫病相关基因。
27.含有至少一种权利要求1-12中任一项的双链RNA分子作为活性剂和一种药用载体的药用组合物。
28.用于诊断的权利要求27的组合物。
29.用于治疗的权利要求27的组合物。
30.一种显示靶基因特异性敲除表型的真核细胞或真核非人类生物,其中用至少一种能够抑制内源靶基因表达的双链RNA分子或用编码能够抑制至少一种内源靶基因表达的至少一种双链RNA分子的DNA转染该细胞或生物。
31.权利要求30的细胞或生物,它是一种哺乳动物细胞。
32.权利要求31的细胞或生物,它是一种人类细胞。
33.权利要求30-32中任一项的细胞或生物,其用至少一种编码靶蛋白或靶蛋白变体或突变形式的外源靶核酸进一步转染,其中该外源靶核酸在核酸水平上不同于内源靶基因,因此与内源靶基因的表达相比,外源靶核酸的表达受双链RNA分子的抑制大大减少。
34.权利要求33的细胞或生物,其中外源靶核酸与编码可检测肽或多肽的另一种核酸序列融合。
35.权利要求30-34中任一项的细胞或生物在分析方法中的用途。
36.权利要求35在基因表达谱分析中的用途。
37.权利要求35在蛋白质组分析中的用途。
38.权利要求35-37中任一项的用途,其中对外源靶核酸编码的靶蛋白的变体或突变形式进行分析。
39.权利要求38在鉴定靶蛋白功能域中的用途。
40.权利要求35-39中任一项的用途,其中对至少两种细胞或生物进行比较,它们选自:
(i)一种对照细胞或对照生物,没有靶基因抑制,
(ii)一种细胞或生物,具有靶基因抑制,和
(iii)一种细胞或生物,具有靶基因抑制加外源靶核酸对靶基因的互补。
41.权利要求35-40中任一项的用途,其中的分析包括功能和/或表型分析。
42.权利要求30-34中任一项的细胞在制备方法中的用途。
43.权利要求41的用途,用于从真核细胞中分离蛋白质或蛋白质复合物。
44.权利要求43的用途,用于分离任选地含有核酸的高分子量蛋白质复合物。
45.权利要求35-44中任一项在鉴定和/或表征药理试剂中的用途。
46.一种系统,用于鉴定和/或表征作用于至少一种靶蛋白的药理试剂,其包括:
(a)一种真核细胞或真核非人类生物,它能表达至少一种编码所述至少一种靶蛋白的内源靶基因,
(b)至少一种双链RNA分子,它能抑制所述至少一种内源靶基因的表达,和
(c)一种待测物质或一组待测物质,其中将要鉴定和/或表征这种待测物质或组合的药理学性质。
47.权利要求46的系统,其进一步包括:
(d)编码靶蛋白或靶蛋白变体或突变形式的至少一种外源靶核酸,其中外源靶核酸在核酸水平上不同于内源靶基因,因此与内源靶基因的表达相比,外源靶核酸的表达受双链RNA分子的抑制大大减少。
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Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100445381C (zh) * 2005-06-10 2008-12-24 中国人民解放军军事医学科学院基础医学研究所 带有单链polyA尾巴的siRNA分子制备方法和应用
CN102333869A (zh) * 2009-02-24 2012-01-25 里博克斯艾克斯有限公司 小干扰rna的改进设计
CN101213300B (zh) * 2005-06-01 2013-01-23 聚加转染股份有限公司 用于rna干扰的寡核苷酸及其生物学应用
US8664189B2 (en) 2008-09-22 2014-03-04 Rxi Pharmaceuticals Corporation RNA interference in skin indications
US8815818B2 (en) 2008-07-18 2014-08-26 Rxi Pharmaceuticals Corporation Phagocytic cell delivery of RNAI
CN104120127A (zh) * 2014-07-01 2014-10-29 清华大学 分离的寡核苷酸及其应用
US9074211B2 (en) 2008-11-19 2015-07-07 Rxi Pharmaceuticals Corporation Inhibition of MAP4K4 through RNAI
US9080171B2 (en) 2010-03-24 2015-07-14 RXi Parmaceuticals Corporation Reduced size self-delivering RNAi compounds
US9095504B2 (en) 2010-03-24 2015-08-04 Rxi Pharmaceuticals Corporation RNA interference in ocular indications
US9340786B2 (en) 2010-03-24 2016-05-17 Rxi Pharmaceuticals Corporation RNA interference in dermal and fibrotic indications
US9493774B2 (en) 2009-01-05 2016-11-15 Rxi Pharmaceuticals Corporation Inhibition of PCSK9 through RNAi
US9745574B2 (en) 2009-02-04 2017-08-29 Rxi Pharmaceuticals Corporation RNA duplexes with single stranded phosphorothioate nucleotide regions for additional functionality
US10131904B2 (en) 2008-02-11 2018-11-20 Rxi Pharmaceuticals Corporation Modified RNAi polynucleotides and uses thereof
US10808247B2 (en) 2015-07-06 2020-10-20 Phio Pharmaceuticals Corp. Methods for treating neurological disorders using a synergistic small molecule and nucleic acids therapeutic approach
US10900039B2 (en) 2014-09-05 2021-01-26 Phio Pharmaceuticals Corp. Methods for treating aging and skin disorders using nucleic acids targeting Tyr or MMP1
US10934550B2 (en) 2013-12-02 2021-03-02 Phio Pharmaceuticals Corp. Immunotherapy of cancer
US11001845B2 (en) 2015-07-06 2021-05-11 Phio Pharmaceuticals Corp. Nucleic acid molecules targeting superoxide dismutase 1 (SOD1)
US11021707B2 (en) 2015-10-19 2021-06-01 Phio Pharmaceuticals Corp. Reduced size self-delivering nucleic acid compounds targeting long non-coding RNA
US11279934B2 (en) 2014-04-28 2022-03-22 Phio Pharmaceuticals Corp. Methods for treating cancer using nucleic acids targeting MDM2 or MYCN

Families Citing this family (1177)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08500481A (ja) * 1992-05-11 1996-01-23 リボザイム・ファーマシューティカルズ・インコーポレーテッド ウイルスの複製を阻害するための方法および薬剤
US20030206887A1 (en) * 1992-05-14 2003-11-06 David Morrissey RNA interference mediated inhibition of hepatitis B virus (HBV) using short interfering nucleic acid (siNA)
US5639647A (en) * 1994-03-29 1997-06-17 Ribozyme Pharmaceuticals, Inc. 2'-deoxy-2'alkylnucleotide containing nucleic acid
US7812149B2 (en) 1996-06-06 2010-10-12 Isis Pharmaceuticals, Inc. 2′-Fluoro substituted oligomeric compounds and compositions for use in gene modulations
US5898031A (en) 1996-06-06 1999-04-27 Isis Pharmaceuticals, Inc. Oligoribonucleotides for cleaving RNA
US9096636B2 (en) 1996-06-06 2015-08-04 Isis Pharmaceuticals, Inc. Chimeric oligomeric compounds and their use in gene modulation
US20040219569A1 (en) * 1999-07-06 2004-11-04 Fruma Yehiely Gene identification method
US20110003879A1 (en) * 2005-03-11 2011-01-06 Vincent Mark D Antisense oligonucleotides targeted to the coding region of thymidylate synthase and uses thereof
AUPP249298A0 (en) 1998-03-20 1998-04-23 Ag-Gene Australia Limited Synthetic genes and genetic constructs comprising same I
WO1999049029A1 (en) 1998-03-20 1999-09-30 Benitec Australia Ltd Control of gene expression
EP1071753A2 (en) * 1998-04-20 2001-01-31 Ribozyme Pharmaceuticals, Inc. Nucleic acid molecules with novel chemical compositions capable of modulating gene expression
CA2361201A1 (en) 1999-01-28 2000-08-03 Medical College Of Georgia Research Institute, Inc. Composition and method for in vivo and in vitro attenuation of gene expression using double stranded rna
DE19956568A1 (de) 1999-01-30 2000-08-17 Roland Kreutzer Verfahren und Medikament zur Hemmung der Expression eines vorgegebenen Gens
US7601494B2 (en) 1999-03-17 2009-10-13 The University Of North Carolina At Chapel Hill Method of screening candidate compounds for susceptibility to biliary excretion
ES2568898T3 (es) 1999-04-09 2016-05-05 Kyowa Hakko Kirin Co., Ltd. Procedimiento para controlar la actividad de una molécula inmunofuncional
US6656698B1 (en) * 1999-06-30 2003-12-02 Millennium Pharmaceuticals, Inc. 12832, a novel human kinase-like molecule and uses thereof
US6423885B1 (en) 1999-08-13 2002-07-23 Commonwealth Scientific And Industrial Research Organization (Csiro) Methods for obtaining modified phenotypes in plant cells
US8128922B2 (en) * 1999-10-20 2012-03-06 Johns Hopkins University Superior molecular vaccine linking the translocation domain of a bacterial toxin to an antigen
GB9925459D0 (en) 1999-10-27 1999-12-29 Plant Bioscience Ltd Gene silencing
US7829693B2 (en) * 1999-11-24 2010-11-09 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of a target gene
DE10100586C1 (de) 2001-01-09 2002-04-11 Ribopharma Ag Verfahren zur Hemmung der Expression eines Ziegens
DE10160151A1 (de) * 2001-01-09 2003-06-26 Ribopharma Ag Verfahren zur Hemmung der Expression eines vorgegebenen Zielgens
US7179796B2 (en) 2000-01-18 2007-02-20 Isis Pharmaceuticals, Inc. Antisense modulation of PTP1B expression
US20050032733A1 (en) * 2001-05-18 2005-02-10 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (SiNA)
WO2005019453A2 (en) * 2001-05-18 2005-03-03 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF GENE EXPRESSION USING CHEMICALLY MODIFIED SHORT INTERFERING NUCLEIC ACID (siNA)
US20070026394A1 (en) * 2000-02-11 2007-02-01 Lawrence Blatt Modulation of gene expression associated with inflammation proliferation and neurite outgrowth using nucleic acid based technologies
US20080039414A1 (en) * 2002-02-20 2008-02-14 Sima Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
US8202979B2 (en) * 2002-02-20 2012-06-19 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid
US8273866B2 (en) 2002-02-20 2012-09-25 Merck Sharp & Dohme Corp. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (SINA)
US20030084471A1 (en) * 2000-03-16 2003-05-01 David Beach Methods and compositions for RNA interference
US8202846B2 (en) 2000-03-16 2012-06-19 Cold Spring Harbor Laboratory Methods and compositions for RNA interference
AU2001245793A1 (en) * 2000-03-16 2001-09-24 Cold Spring Harbor Laboratory Methods and compositions for rna interference
EP2796553B1 (en) * 2000-03-30 2019-06-19 Whitehead Institute for Biomedical Research RNA sequence-specific mediators of RNA interference
AU2001249622B2 (en) 2000-03-30 2007-06-07 Massachusetts Institute Of Technology RNA sequence-specific mediators of RNA interference
US7662791B2 (en) * 2000-08-02 2010-02-16 University Of Southern California Gene silencing using mRNA-cDNA hybrids
US20080242627A1 (en) * 2000-08-02 2008-10-02 University Of Southern California Novel rna interference methods using dna-rna duplex constructs
JP5087201B2 (ja) * 2000-08-03 2012-12-05 ジョンズ・ホプキンス・ユニバーシティ 抗原に小胞体シャペロンポリペプチドを連結した分子ワクチン
US20030190635A1 (en) * 2002-02-20 2003-10-09 Mcswiggen James A. RNA interference mediated treatment of Alzheimer's disease using short interfering RNA
US20080032942A1 (en) 2000-08-30 2008-02-07 Mcswiggen James RNA interference mediated treatment of Alzheimer's disease using short interfering nucleic acid (siNA)
WO2009042910A2 (en) * 2007-09-26 2009-04-02 University Of South Florida Ship inhibition to direct hematopoietic stem cells and induce extramedullary hematopoiesis
US7691821B2 (en) 2001-09-19 2010-04-06 University Of South Florida Inhibition of SHIP to enhance stem cell harvest and transplantation
US20020165192A1 (en) * 2000-09-19 2002-11-07 Kerr William G. Control of NK cell function and survival by modulation of ship activity
US6946292B2 (en) 2000-10-06 2005-09-20 Kyowa Hakko Kogyo Co., Ltd. Cells producing antibody compositions with increased antibody dependent cytotoxic activity
ES2728168T3 (es) 2000-12-01 2019-10-22 Max Planck Gesellschaft Moléculas pequeñas de ARN que median en la interferencia de ARN
US8546143B2 (en) 2001-01-09 2013-10-01 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of a target gene
US7767802B2 (en) 2001-01-09 2010-08-03 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of anti-apoptotic genes
CA2369944A1 (en) * 2001-01-31 2002-07-31 Nucleonics Inc. Use of post-transcriptional gene silencing for identifying nucleic acid sequences that modulate the function of a cell
US20050176663A1 (en) * 2001-05-18 2005-08-11 Sima Therapeutics, Inc. RNA interference mediated inhibition of protein tyrosine phosphatase type IVA (PRL3) gene expression using short interfering nucleic acid (siNA)
US20050187174A1 (en) * 2001-05-18 2005-08-25 Sirna Therapeutics, Inc. RNA interference mediated inhibition of intercellular adhesion molecule (ICAM) gene expression using short interfering nucleic acid (siNA)
US20050203040A1 (en) * 2001-05-18 2005-09-15 Sirna Therapeutics, Inc. RNA interference mediated inhibition of vascular cell adhesion molecule (VCAM) gene expression using short interfering nucleic acid (siNA)
US20050233997A1 (en) * 2001-05-18 2005-10-20 Sirna Therapeutics, Inc. RNA interference mediated inhibition of matrix metalloproteinase 13 (MMP13) gene expression using short interfering nucleic acid (siNA)
US20050159378A1 (en) * 2001-05-18 2005-07-21 Sirna Therapeutics, Inc. RNA interference mediated inhibition of Myc and/or Myb gene expression using short interfering nucleic acid (siNA)
US20050148530A1 (en) 2002-02-20 2005-07-07 Sirna Therapeutics, Inc. RNA interference mediated inhibition of vascular endothelial growth factor and vascular endothelial growth factor receptor gene expression using short interfering nucleic acid (siNA)
US9994853B2 (en) 2001-05-18 2018-06-12 Sirna Therapeutics, Inc. Chemically modified multifunctional short interfering nucleic acid molecules that mediate RNA interference
US20050196767A1 (en) * 2001-05-18 2005-09-08 Sirna Therapeutics, Inc. RNA interference mediated inhibition of GRB2 associated binding protein (GAB2) gene expression using short interfering nucleic acis (siNA)
US20050287128A1 (en) * 2001-05-18 2005-12-29 Sirna Therapeutics, Inc. RNA interference mediated inhibition of TGF-beta and TGF-beta receptor gene expression using short interfering nucleic acid (siNA)
US20050119212A1 (en) * 2001-05-18 2005-06-02 Sirna Therapeutics, Inc. RNA interference mediated inhibition of FAS and FASL gene expression using short interfering nucleic acid (siNA)
US20050196765A1 (en) * 2001-05-18 2005-09-08 Sirna Therapeutics, Inc. RNA interference mediated inhibition of checkpoint Kinase-1 (CHK-1) gene expression using short interfering nucleic acid (siNA)
US20050048529A1 (en) * 2002-02-20 2005-03-03 Sirna Therapeutics, Inc. RNA interference mediated inhibition of intercellular adhesion molecule (ICAM) gene expression using short interfering nucleic acid (siNA)
US20050014172A1 (en) 2002-02-20 2005-01-20 Ivan Richards RNA interference mediated inhibition of muscarinic cholinergic receptor gene expression using short interfering nucleic acid (siNA)
WO2005014811A2 (en) * 2003-08-08 2005-02-17 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF XIAP GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
US20050159382A1 (en) * 2001-05-18 2005-07-21 Sirna Therapeutics, Inc. RNA interference mediated inhibition of polycomb group protein EZH2 gene expression using short interfering nucleic acid (siNA)
US20050176666A1 (en) * 2001-05-18 2005-08-11 Sirna Therapeutics, Inc. RNA interference mediated inhibition of GPRA and AAA1 gene expression using short interfering nucleic acid (siNA)
US20050159380A1 (en) * 2001-05-18 2005-07-21 Sirna Therapeutics, Inc. RNA interference mediated inhibition of angiopoietin gene expression using short interfering nucleic acid (siNA)
US7109165B2 (en) * 2001-05-18 2006-09-19 Sirna Therapeutics, Inc. Conjugates and compositions for cellular delivery
US20060148743A1 (en) * 2001-05-18 2006-07-06 Vasant Jadhav RNA interference mediated inhibition of histone deacetylase (HDAC) gene expression using short interfering nucleic acid (siNA)
US20040219671A1 (en) * 2002-02-20 2004-11-04 Sirna Therapeutics, Inc. RNA interference mediated treatment of parkinson disease using short interfering nucleic acid (siNA)
US20070093437A1 (en) * 2001-05-18 2007-04-26 Sirna Therapeutics, Inc. Rna interference mediated inhibition of xiap gene expression using short interfering nucleic acid (sina)
US20050233996A1 (en) * 2002-02-20 2005-10-20 Sirna Therapeutics, Inc. RNA interference mediated inhibition of hairless (HR) gene expression using short interfering nucleic acid (siNA)
US20050176664A1 (en) * 2001-05-18 2005-08-11 Sirna Therapeutics, Inc. RNA interference mediated inhibition of cholinergic muscarinic receptor (CHRM3) gene expression using short interfering nucleic acid (siNA)
US20050159379A1 (en) * 2001-05-18 2005-07-21 Sirna Therapeutics, Inc RNA interference mediated inhibition of gastric inhibitory polypeptide (GIP) and gastric inhibitory polypeptide receptor (GIPR) gene expression using short interfering nucleic acid (siNA)
US20050158735A1 (en) * 2001-05-18 2005-07-21 Sirna Therapeutics, Inc. RNA interference mediated inhibition of proliferating cell nuclear antigen (PCNA) gene expression using short interfering nucleic acid (siNA)
US20060211642A1 (en) * 2001-05-18 2006-09-21 Sirna Therapeutics, Inc. RNA inteference mediated inhibition of hepatitis C virus (HVC) gene expression using short interfering nucleic acid (siNA)
US20070270579A1 (en) * 2001-05-18 2007-11-22 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using short interfering nucleic acid (siNA)
US20080188430A1 (en) * 2001-05-18 2008-08-07 Sirna Therapeutics, Inc. RNA interference mediated inhibition of hypoxia inducible factor 1 (HIF1) gene expression using short interfering nucleic acid (siNA)
US20050153914A1 (en) * 2001-05-18 2005-07-14 Sirna Therapeutics, Inc. RNA interference mediated inhibition of MDR P-glycoprotein gene expression using short interfering nucleic acid (siNA)
US20040198682A1 (en) * 2001-11-30 2004-10-07 Mcswiggen James RNA interference mediated inhibition of placental growth factor gene expression using short interfering nucleic acid (siNA)
US7517864B2 (en) 2001-05-18 2009-04-14 Sirna Therapeutics, Inc. RNA interference mediated inhibition of vascular endothelial growth factor and vascular endothelial growth factor receptor gene expression using short interfering nucleic acid (siNA)
US20070042983A1 (en) * 2001-05-18 2007-02-22 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using short interfering nucleic acid (siNA)
US20050124569A1 (en) * 2001-05-18 2005-06-09 Sirna Therapeutics, Inc. RNA interference mediated inhibition of CXCR4 gene expression using short interfering nucleic acid (siNA)
US20050196781A1 (en) * 2001-05-18 2005-09-08 Sirna Therapeutics, Inc. RNA interference mediated inhibition of STAT3 gene expression using short interfering nucleic acid (siNA)
US20050124566A1 (en) * 2001-05-18 2005-06-09 Sirna Therapeutics, Inc. RNA interference mediated inhibition of myostatin gene expression using short interfering nucleic acid (siNA)
US20050137155A1 (en) * 2001-05-18 2005-06-23 Sirna Therapeutics, Inc. RNA interference mediated treatment of Parkinson disease using short interfering nucleic acid (siNA)
US20030175950A1 (en) * 2001-05-29 2003-09-18 Mcswiggen James A. RNA interference mediated inhibition of HIV gene expression using short interfering RNA
US20050159381A1 (en) * 2001-05-18 2005-07-21 Sirna Therapeutics, Inc. RNA interference mediated inhibition of chromosome translocation gene expression using short interfering nucleic acid (siNA)
US20050288242A1 (en) * 2001-05-18 2005-12-29 Sirna Therapeutics, Inc. RNA interference mediated inhibition of RAS gene expression using short interfering nucleic acid (siNA)
US20050164967A1 (en) * 2001-05-18 2005-07-28 Sirna Therapeutics, Inc. RNA interference mediated inhibition of platelet-derived endothelial cell growth factor (ECGF1) gene expression using short interfering nucleic acid (siNA)
US20040019001A1 (en) * 2002-02-20 2004-01-29 Mcswiggen James A. RNA interference mediated inhibition of protein typrosine phosphatase-1B (PTP-1B) gene expression using short interfering RNA
US20050176025A1 (en) * 2001-05-18 2005-08-11 Sirna Therapeutics, Inc. RNA interference mediated inhibition of B-cell CLL/Lymphoma-2 (BCL-2) gene expression using short interfering nucleic acid (siNA)
US20050164224A1 (en) * 2001-05-18 2005-07-28 Sirna Therapeutics, Inc. RNA interference mediated inhibition of cyclin D1 gene expression using short interfering nucleic acid (siNA)
US20050079610A1 (en) * 2001-05-18 2005-04-14 Sirna Therapeutics, Inc. RNA interference mediated inhibition of Fos gene expression using short interfering nucleic acid (siNA)
US20050182007A1 (en) * 2001-05-18 2005-08-18 Sirna Therapeutics, Inc. RNA interference mediated inhibition of interleukin and interleukin receptor gene expression using short interfering nucleic acid (SINA)
US20050136436A1 (en) * 2001-05-18 2005-06-23 Sirna Therapeutics, Inc. RNA interference mediated inhibition of G72 and D-amino acid oxidase (DAAO) gene expression using short interfering nucleic acid (siNA)
US20090299045A1 (en) * 2001-05-18 2009-12-03 Sirna Therapeutics, Inc. RNA Interference Mediated Inhibition Of Interleukin and Interleukin Gene Expression Using Short Interfering Nucleic Acid (siNA)
US20050239731A1 (en) * 2001-05-18 2005-10-27 Sirna Therapeutics, Inc. RNA interference mediated inhibition of MAP kinase gene expression using short interfering nucleic acid (siNA)
US20060142225A1 (en) * 2001-05-18 2006-06-29 Sirna Therapeutics, Inc. RNA interference mediated inhibition of cyclin dependent kinase-2 (CDK2) gene expression using short interfering nucleic acid (siNA)
US20050261219A1 (en) * 2001-05-18 2005-11-24 Sirna Therapeutics, Inc. RNA interference mediated inhibition of interleukin and interleukin receptor gene expression using short interfering nucleic acid (siNA)
US20080161256A1 (en) * 2001-05-18 2008-07-03 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using short interfering nucleic acid (siNA)
WO2003070983A1 (en) * 2002-02-20 2003-08-28 Sirna Therapeutics, Inc RNA INTERFERENCE MEDIATED INHIBITION OF PROTEIN KINASE C ALPHA (PKC-ALPHA) GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
US20050256068A1 (en) 2001-05-18 2005-11-17 Sirna Therapeutics, Inc. RNA interference mediated inhibition of stearoyl-CoA desaturase (SCD) gene expression using short interfering nucleic acid (siNA)
US20050143333A1 (en) * 2001-05-18 2005-06-30 Sirna Therapeutics, Inc. RNA interference mediated inhibition of interleukin and interleukin receptor gene expression using short interfering nucleic acid (SINA)
US20050164968A1 (en) * 2001-05-18 2005-07-28 Sirna Therapeutics, Inc. RNA interference mediated inhibition of ADAM33 gene expression using short interfering nucleic acid (siNA)
US20050267058A1 (en) * 2001-05-18 2005-12-01 Sirna Therapeutics, Inc. RNA interference mediated inhibition of placental growth factor gene expression using short interfering nucleic acid (sINA)
US20050282188A1 (en) * 2001-05-18 2005-12-22 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using short interfering nucleic acid (siNA)
US20050191618A1 (en) * 2001-05-18 2005-09-01 Sirna Therapeutics, Inc. RNA interference mediated inhibition of human immunodeficiency virus (HIV) gene expression using short interfering nucleic acid (siNA)
US20050054596A1 (en) * 2001-11-30 2005-03-10 Mcswiggen James RNA interference mediated inhibition of vascular endothelial growth factor and vascular endothelial growth factor receptor gene expression using short interfering nucleic acid (siNA)
US20050233344A1 (en) * 2001-05-18 2005-10-20 Sirna Therapeutics, Inc. RNA interference mediated inhibition of platelet derived growth factor (PDGF) and platelet derived growth factor receptor (PDGFR) gene expression using short interfering nucleic acid (siNA)
US20050209180A1 (en) * 2001-05-18 2005-09-22 Sirna Therapeutics, Inc. RNA interference mediated inhibition of hepatitis C virus (HCV) expression using short interfering nucleic acid (siNA)
US20050176024A1 (en) * 2001-05-18 2005-08-11 Sirna Therapeutics, Inc. RNA interference mediated inhibition of epidermal growth factor receptor (EGFR) gene expression using short interfering nucleic acid (siNA)
US20050222066A1 (en) * 2001-05-18 2005-10-06 Sirna Therapeutics, Inc. RNA interference mediated inhibition of vascular endothelial growth factor and vascular endothelial growth factor receptor gene expression using short interfering nucleic acid (siNA)
US8008472B2 (en) 2001-05-29 2011-08-30 Merck Sharp & Dohme Corp. RNA interference mediated inhibition of human immunodeficiency virus (HIV) gene expression using short interfering nucleic acid (siNA)
EP1390472A4 (en) * 2001-05-29 2004-11-17 Sirna Therapeutics Inc NUCLEIC ACID TREATMENT OF DISEASES OR SIDES RELATED TO RAS, HER2 AND HIV LEVELS
US20050019915A1 (en) 2001-06-21 2005-01-27 Bennett C. Frank Antisense modulation of superoxide dismutase 1, soluble expression
EP1404698A4 (en) 2001-06-21 2004-12-22 Isis Pharmaceuticals Inc ANTI-SENSE MODULATION OF SOLUBLE SUPEROXIDE DISMUTASE 1 EXPRESSION
DE10133858A1 (de) * 2001-07-12 2003-02-06 Aventis Pharma Gmbh Synthetische doppelsträngige Oligonucleotide zur gezielten Hemmung der Genexpression
US7691995B2 (en) 2001-07-12 2010-04-06 University Of Massachusetts In vivo production of small interfering RNAS that mediate gene silencing
US10590418B2 (en) 2001-07-23 2020-03-17 The Board Of Trustees Of The Leland Stanford Junior University Methods and compositions for RNAi mediated inhibition of gene expression in mammals
EP1409506B1 (en) * 2001-07-23 2012-05-09 The Board Of Trustees Of The Leland Stanford Junior University Methods and compositions for rnai mediated inhibition of gene expression in mammals
US20090247606A1 (en) * 2001-08-28 2009-10-01 Sirna Therapeutics, Inc. RNA Interference Mediated Inhibition of Adenosine A1 Receptor (ADORA1) Gene Expression Using Short Interfering Nucleic Acid (siNA)
US20030198627A1 (en) * 2001-09-01 2003-10-23 Gert-Jan Arts siRNA knockout assay method and constructs
DE10163098B4 (de) 2001-10-12 2005-06-02 Alnylam Europe Ag Verfahren zur Hemmung der Replikation von Viren
US7745418B2 (en) 2001-10-12 2010-06-29 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting viral replication
CN1604783A (zh) * 2001-10-26 2005-04-06 里伯药品公司 通过rna干扰治疗纤维化疾病的药物
WO2003035870A1 (de) * 2001-10-26 2003-05-01 Ribopharma Ag Medikament zur behandlung eines pankreaskarzinoms
DE10230996A1 (de) * 2001-10-26 2003-07-17 Ribopharma Ag Medikament zur Behandlung eines Pankreaskarzinoms
US20040248835A1 (en) * 2001-10-26 2004-12-09 Anja Krebs Use of a double-stranded ribonucleic acid for treating an infection with a positivestrand rna-virus
WO2003040399A2 (en) * 2001-11-02 2003-05-15 Intradigm Corporation Therapeutic methods for nucleic acid delivery vehicles
US20040063654A1 (en) * 2001-11-02 2004-04-01 Davis Mark E. Methods and compositions for therapeutic use of RNA interference
CA2465860A1 (en) * 2001-11-02 2004-04-22 Insert Therapeutics, Inc. Methods and compositions for therapeutic use of rna interference
EP1445312B1 (en) * 2001-11-21 2012-12-26 Astellas Pharma Inc. Method of inhibiting gene expression
US20040138163A1 (en) * 2002-05-29 2004-07-15 Mcswiggen James RNA interference mediated inhibition of vascular edothelial growth factor and vascular edothelial growth factor receptor gene expression using short interfering nucleic acid (siNA)
US20050075304A1 (en) * 2001-11-30 2005-04-07 Mcswiggen James RNA interference mediated inhibition of vascular endothelial growth factor and vascular endothelial growth factor receptor gene expression using short interfering nucleic acid (siNA)
US20070203333A1 (en) * 2001-11-30 2007-08-30 Mcswiggen James RNA interference mediated inhibition of vascular endothelial growth factor and vascular endothelial growth factor receptor gene expression using short interfering nucleic acid (siNA)
US7294504B1 (en) 2001-12-27 2007-11-13 Allele Biotechnology & Pharmaceuticals, Inc. Methods and compositions for DNA mediated gene silencing
NZ533126A (en) * 2002-01-17 2006-04-28 Univ British Columbia Bispecific antisense oligonucleotides that inhibit IGFBP-2 and IGFBP-5 and methods of using same
DE10202419A1 (de) 2002-01-22 2003-08-07 Ribopharma Ag Verfahren zur Hemmung der Expression eines durch eine Chromosomen-Aberration entstandenen Zielgens
GB0201477D0 (en) * 2002-01-23 2002-03-13 Novartis Forschungsstiftung Methods of obtaining isoform specific expression in mammalian cells
US20060009409A1 (en) * 2002-02-01 2006-01-12 Woolf Tod M Double-stranded oligonucleotides
WO2003064621A2 (en) * 2002-02-01 2003-08-07 Ambion, Inc. HIGH POTENCY siRNAS FOR REDUCING THE EXPRESSION OF TARGET GENES
EP1478656B1 (en) 2002-02-01 2009-09-16 Life Technologies Corporation Oligonucleotide compositions with enhanced efficiency
US20050096289A1 (en) * 2002-02-07 2005-05-05 Hans Prydz Methods and compositions for modulating tissue factor
EP1483400A4 (en) * 2002-02-12 2007-07-11 Quark Biotech Inc USE OF THE AXL RECEPTOR FOR THE DIAGNOSIS AND TREATMENT OF A KIDNEY DISEASE
EP1474512A2 (en) * 2002-02-13 2004-11-10 Axordia Limited Method to modify differentiation of pluripotential stem cells
JP2005527198A (ja) 2002-02-14 2005-09-15 シティ・オブ・ホープ 哺乳動物細胞において、干渉rna分子を産生する方法、および該干渉rna分子の療法的使用
US20090247613A1 (en) * 2002-02-20 2009-10-01 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF B-CELL CLL/LYMPHOMA-2 (BCL2) GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
US7667030B2 (en) 2002-02-20 2010-02-23 Sirna Therapeutics, Inc. RNA interference mediated inhibition of matrix metalloproteinase 13 (MMP13) gene expression using short interfering nucleic acid (siNA)
US20050096284A1 (en) * 2002-02-20 2005-05-05 Sirna Therapeutics, Inc. RNA interference mediated treatment of polyglutamine (polyQ) repeat expansion diseases using short interfering nucleic acid (siNA)
US7667029B2 (en) 2002-02-20 2010-02-23 Sirna Therapeutics, Inc. RNA interference mediated inhibition of checkpoint kinase-1 (CHK-1) gene expression using short interfering nucleic acid (siNA)
US7700760B2 (en) 2002-02-20 2010-04-20 Sirna Therapeutics, Inc. RNA interference mediated inhibition of vascular cell adhesion molecule (VCAM) gene expression using short interfering nucleic acid (siNA)
US8258288B2 (en) 2002-02-20 2012-09-04 Sirna Therapeutics, Inc. RNA interference mediated inhibition of respiratory syncytial virus (RSV) expression using short interfering nucleic acid (siNA)
US20090233983A1 (en) * 2002-02-20 2009-09-17 Sirna Therapeutics Inc. RNA Interference Mediated Inhibition of Protein Tyrosine Phosphatase-1B (PTP-1B) Gene Expression Using Short Interfering RNA
US7691999B2 (en) 2002-02-20 2010-04-06 Sirna Therapeutics, Inc. RNA interference mediated inhibition of NOGO and NOGO receptor gene expression using short interfering nucleic acid (siNA)
US7928219B2 (en) 2002-02-20 2011-04-19 Merck Sharp & Dohme Corp. RNA interference mediated inhibition of placental growth factor gene expression using short interfering nucleic acid (SINA)
EP1432724A4 (en) 2002-02-20 2006-02-01 Sirna Therapeutics Inc RNA inhibition mediated inhibition of MAP KINASE GENES
US7897757B2 (en) 2002-02-20 2011-03-01 Merck Sharp & Dohme Corp. RNA interference mediated inhibition of protein tyrosine phosphatase-1B (PTP-1B) gene expression using short interfering nucleic acid (siNA)
JP2005517433A (ja) * 2002-02-20 2005-06-16 サーナ・セラピューティクス・インコーポレイテッド 短干渉核酸(siNA)を用いるTNFおよびTNFレセプタースーパーファミリー遺伝子発現のRNA干渉媒介性阻害
US9657294B2 (en) 2002-02-20 2017-05-23 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
US7678897B2 (en) * 2002-02-20 2010-03-16 Sirna Therapeutics, Inc. RNA interference mediated inhibition of platelet-derived endothelial cell growth factor (ECGF1) gene expression using short interfering nucleic acid (siNA)
US20090137509A1 (en) * 2002-02-20 2009-05-28 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF PROLIFERATION CELL NUCLEAR ANTIGEN (PCNA) GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
US8067575B2 (en) 2002-02-20 2011-11-29 Merck, Sharp & Dohme Corp. RNA interference mediated inhibition of cyclin D1 gene expression using short interfering nucleic acid (siNA)
US7662952B2 (en) 2002-02-20 2010-02-16 Sirna Therapeutics, Inc. RNA interference mediated inhibition of GRB2 associated binding protein (GAB2) gene expression using short interfering nucleic acid (siNA)
EP1495041A4 (en) * 2002-02-20 2006-02-01 Sirna Therapeutics Inc RNA interferon-mediated inhibition of gene expression of G72 and D-amino acid oxidase (DAAO) using short-term interfering nucleic acid (siNA)
US7683166B2 (en) 2002-02-20 2010-03-23 Sirna Therapeutics, Inc. RNA interference mediated inhibition of interleukin and interleukin receptor gene expression using short interfering nucleic acid (siNA)
US8232383B2 (en) * 2002-02-20 2012-07-31 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
US20090306182A1 (en) * 2002-02-20 2009-12-10 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF MAP KINASE GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
US20090093439A1 (en) * 2002-02-20 2009-04-09 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF CHROMOSOME TRANSLOCATION GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
US20100240730A1 (en) * 2002-02-20 2010-09-23 Merck Sharp And Dohme Corp. RNA Interference Mediated Inhibition of Gene Expression Using Chemically Modified Short Interfering Nucleic Acid (siNA)
US9181551B2 (en) 2002-02-20 2015-11-10 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
US7928220B2 (en) 2002-02-20 2011-04-19 Merck Sharp & Dohme Corp. RNA interference mediated inhibition of stromal cell-derived factor-1 (SDF-1) gene expression using short interfering nucleic acid (siNA)
US7683165B2 (en) 2002-02-20 2010-03-23 Sirna Therapeutics, Inc. RNA interference mediated inhibition of interleukin and interleukin receptor gene expression using short interfering nucleic acid (siNA)
EP1474433A4 (en) * 2002-02-20 2005-02-23 Sirna Therapeutics Inc TARGET LOCALIZATION TARGETED BY RNA INTERFERENCE AND TARGET VALIDATION WITH SHORT INTERFERING NUCLEIC ACID (siNA)
US7897753B2 (en) 2002-02-20 2011-03-01 Sirna Therapeutics, Inc. RNA interference mediated inhibition of XIAP gene expression using short interfering nucleic acid (siNA)
US20090099117A1 (en) 2002-02-20 2009-04-16 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF MYOSTATIN GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
US7897752B2 (en) 2002-02-20 2011-03-01 Sirna Therapeutics, Inc. RNA interference mediated inhibition of telomerase gene expression using short interfering nucleic acid (siNA)
GB2397062B (en) 2002-02-20 2005-06-15 Sirna Therapeutics Inc RNA interference mediated inhibition of hepatitis c virus (HCV) gene expression using short interfering nucleic acid (siNA)
WO2003106476A1 (en) * 2002-02-20 2003-12-24 Sirna Therapeutics, Inc Nucleic acid mediated inhibition of enterococcus infection and cytolysin toxin activity
US20090253774A1 (en) 2002-02-20 2009-10-08 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF PLATELET DERIVED GROWTH FACTOR (PDGF) AND PLATELET DERIVED GROWTH FACTOR RECEPTOR (PDGFR) GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
EP1499631A4 (en) * 2002-02-20 2006-01-25 Sirna Therapeutics Inc RNA INTERFERENCE-ADMINISTRATIVE INHIBITION OF TGF-BETA AND TGF-BETA-RECEPTOR GENE EXPRESSION WITH SHORT-INTERFERENCE NUCLEIC ACID (SINA)
US20090192105A1 (en) 2002-02-20 2009-07-30 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF INTERCELLULAR ADHESION MOLECULE (ICAM) GENE EXPRESSION USING SHORT INTERFERING NUCELIC ACID (siNA)
US7910724B2 (en) 2002-02-20 2011-03-22 Sirna Therapeutics, Inc. RNA interference mediated inhibition of Fos gene expression using short interfering nucleic acid (siNA)
US20090253773A1 (en) 2002-02-20 2009-10-08 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF TNF AND TNF RECEPTOR GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
US7935812B2 (en) 2002-02-20 2011-05-03 Merck Sharp & Dohme Corp. RNA interference mediated inhibition of hepatitis C virus (HCV) expression using short interfering nucleic acid (siNA)
US7795422B2 (en) 2002-02-20 2010-09-14 Sirna Therapeutics, Inc. RNA interference mediated inhibition of hypoxia inducible factor 1 (HIF1) gene expression using short interfering nucleic acid (siNA)
US7928218B2 (en) 2002-02-20 2011-04-19 Merck Sharp & Dohme Corp. RNA interference mediated inhibition of polycomb group protein EZH2 gene expression using short interfering nucleic acid (siNA)
JP2005517437A (ja) * 2002-02-20 2005-06-16 サーナ・セラピューティクス・インコーポレイテッド 短干渉核酸(siNa)を用いる表皮成長因子レセプター遺伝子発現のRNA干渉媒介性阻害
US8013143B2 (en) 2002-02-20 2011-09-06 Merck Sharp & Dohme Corp. RNA interference mediated inhibition of CXCR4 gene expression using short interfering nucleic acid (siNA)
US7893248B2 (en) 2002-02-20 2011-02-22 Sirna Therapeutics, Inc. RNA interference mediated inhibition of Myc and/or Myb gene expression using short interfering nucleic acid (siNA)
AU2003219900A1 (en) * 2002-02-22 2003-09-09 James R. Eshleman Antigene locks and therapeutic uses thereof
EP1575481A4 (en) * 2002-03-01 2010-01-06 Celltech R & D Inc PROCESS FOR INCREASING OR REDUCING THE BONE DENSITY
US20040005593A1 (en) * 2002-03-06 2004-01-08 Rigel Pharmaceuticals, Inc. Novel method for delivery and intracellular synthesis of siRNA molecules
US7274703B2 (en) * 2002-03-11 2007-09-25 3Com Corporation Stackable network units with resiliency facility
EP1495141A4 (en) * 2002-03-20 2006-03-22 Massachusetts Inst Technology HIV THERAPEUTIC
US7357928B2 (en) 2002-04-08 2008-04-15 University Of Louisville Research Foundation, Inc. Method for the diagnosis and prognosis of malignant diseases
EP1495121A2 (en) * 2002-04-18 2005-01-12 Lynkeus Biotech GmbH Means and methods for the specific inhibition of genes in cells and tissue of the cns and/or eye
PT1504126E (pt) 2002-05-03 2014-06-02 Univ Duke Um método para regular a expressão génica
CA2525976A1 (en) 2002-05-23 2003-12-04 Ceptyr, Inc. Modulation of ptp1b signal transduction by rna interference
US7199107B2 (en) * 2002-05-23 2007-04-03 Isis Pharmaceuticals, Inc. Antisense modulation of kinesin-like 1 expression
AU2003237686A1 (en) * 2002-05-24 2003-12-12 Max-Planck Gesellschaft Zur Forderung Der Wissenschaften E.V. Rna interference mediating small rna molecules
US20100075423A1 (en) * 2002-06-12 2010-03-25 Life Technologies Corporation Methods and compositions relating to polypeptides with rnase iii domains that mediate rna interference
US20040248094A1 (en) * 2002-06-12 2004-12-09 Ford Lance P. Methods and compositions relating to labeled RNA molecules that reduce gene expression
GB2406169B (en) * 2002-06-12 2006-11-01 Ambion Inc Methods and compositions relating to labeled rna molecules that reduce gene expression
EP1513538A4 (en) * 2002-06-14 2007-08-22 Mirus Bio Corp Novel methods for introducing polynucleotides into cells
CN1662652B (zh) * 2002-06-21 2011-05-25 北京诺赛基因组研究中心有限公司 随机化的dna文库和双链rna文库,其用途及生产方法
AU2003256325A1 (en) * 2002-06-26 2004-01-19 The Penn State Research Foundation Methods and materials for treating human papillomavirus infections
US7901708B2 (en) 2002-06-28 2011-03-08 Protiva Biotherapeutics, Inc. Liposomal apparatus and manufacturing methods
EP2333062A1 (en) 2002-07-10 2011-06-15 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. RNA-interference by single-stranded RNA molecules
US7148342B2 (en) 2002-07-24 2006-12-12 The Trustees Of The University Of Pennyslvania Compositions and methods for sirna inhibition of angiogenesis
AU2015264957B2 (en) * 2002-08-05 2017-10-26 Silence Therapeutics Gmbh Further novel forms of interfering rna molecules
WO2004015107A2 (en) 2002-08-05 2004-02-19 Atugen Ag Further novel forms of interfering rna molecules
US20040241854A1 (en) 2002-08-05 2004-12-02 Davidson Beverly L. siRNA-mediated gene silencing
ES2695050T3 (es) * 2002-08-05 2018-12-28 Silence Therapeutics Gmbh Nuevas formas adicionales de moléculas de ARN de interferencia
DK1389637T3 (da) 2002-08-05 2012-09-03 Silence Therapeutics Ag Interfererende RNA-molekyler med stumpe ender
US20080274989A1 (en) 2002-08-05 2008-11-06 University Of Iowa Research Foundation Rna Interference Suppression of Neurodegenerative Diseases and Methods of Use Thereof
US20050042646A1 (en) 2002-08-05 2005-02-24 Davidson Beverly L. RNA interference suppresion of neurodegenerative diseases and methods of use thereof
AU2012216354B2 (en) * 2002-08-05 2016-01-14 Silence Therapeutics Gmbh Further novel forms of interfering RNA molecules
AU2003258100A1 (en) * 2002-08-06 2004-02-23 Intradigm Corporation Methods of down regulating target gene expression in vivo by introduction of interfering rna
WO2004017997A1 (ja) * 2002-08-06 2004-03-04 Toray Industries, Inc. 腎疾患治療又は予防剤及び腎疾患の診断方法
WO2004014933A1 (en) 2002-08-07 2004-02-19 University Of Massachusetts Compositions for rna interference and methods of use thereof
US20040029275A1 (en) * 2002-08-10 2004-02-12 David Brown Methods and compositions for reducing target gene expression using cocktails of siRNAs or constructs expressing siRNAs
DK1536827T3 (da) * 2002-08-14 2009-04-20 Silence Therapeutics Ag Anvendelse af proteinkinase N-beta
KR101238701B1 (ko) 2002-08-21 2013-03-05 더 유니버시티 오브 브리티쉬 콜롬비아 암-관련 단백질을 표적으로 하는 알엔에이아이 프로브
US7956176B2 (en) * 2002-09-05 2011-06-07 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
US20060287269A1 (en) * 2002-09-09 2006-12-21 The Regents Of The University Of California Short interfering nucleic acid hybrids and methods thereof
US20080260744A1 (en) 2002-09-09 2008-10-23 Omeros Corporation G protein coupled receptors and uses thereof
US20040138119A1 (en) * 2002-09-18 2004-07-15 Ingo Tamm Use of hepatitis B X-interacting protein (HBXIP) in modulation of apoptosis
US20060257380A1 (en) * 2002-09-19 2006-11-16 Inst.Nat. De La Sante Et De La Recherche MED Use of sirnas for gene silencing in antigen presenting cells
EP1556402B1 (en) * 2002-09-25 2011-06-22 University of Massachusetts In vivo gene silencing by chemically modified and stable sirna
WO2004028471A2 (en) * 2002-09-28 2004-04-08 Massachusetts Institute Of Technology Influenza therapeutic
US20060160759A1 (en) * 2002-09-28 2006-07-20 Jianzhu Chen Influenza therapeutic
US20040242518A1 (en) * 2002-09-28 2004-12-02 Massachusetts Institute Of Technology Influenza therapeutic
US20060240425A1 (en) * 2002-09-30 2006-10-26 Oncotherapy Science, Inc Genes and polypeptides relating to myeloid leukemia
US7422853B1 (en) * 2002-10-04 2008-09-09 Myriad Genetics, Inc. RNA interference using a universal target
WO2004042024A2 (en) * 2002-11-01 2004-05-21 The Trustees Of The University Of Pennsylvania COMPOSITIONS AND METHODS FOR siRNA INHIBITION OF HIF-1 ALPHA
US7892793B2 (en) 2002-11-04 2011-02-22 University Of Massachusetts Allele-specific RNA interference
US9150606B2 (en) 2002-11-05 2015-10-06 Isis Pharmaceuticals, Inc. Compositions comprising alternating 2'-modified nucleosides for use in gene modulation
AU2003291682A1 (en) * 2002-11-05 2004-06-03 Isis Pharmaceuticals, Inc. 2'-methoxy substituted oligomeric compounds and compositions for use in gene modulations
US9150605B2 (en) 2002-11-05 2015-10-06 Isis Pharmaceuticals, Inc. Compositions comprising alternating 2′-modified nucleosides for use in gene modulation
WO2004044139A2 (en) 2002-11-05 2004-05-27 Isis Parmaceuticals, Inc. Modified oligonucleotides for use in rna interference
WO2004041889A2 (en) 2002-11-05 2004-05-21 Isis Pharmaceuticals, Inc. Polycyclic sugar surrogate-containing oligomeric compounds and compositions for use in gene modulation
DE10322662A1 (de) * 2002-11-06 2004-10-07 Grünenthal GmbH Wirksame und stabile DNA-Enzyme
US10011836B2 (en) 2002-11-14 2018-07-03 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US7655785B1 (en) 2002-11-14 2010-02-02 Rosetta Genomics Ltd. Bioinformatically detectable group of novel regulatory oligonucleotides and uses thereof
US7612196B2 (en) 2002-11-14 2009-11-03 Dharmacon, Inc. siRNA targeting cyclin-dependent kinase inhibitor 1B (p27, Kip1) (CDKN1B)
US7691998B2 (en) * 2002-11-14 2010-04-06 Dharmacon, Inc. siRNA targeting nucleoporin 62kDa (Nup62)
US7250496B2 (en) 2002-11-14 2007-07-31 Rosetta Genomics Ltd. Bioinformatically detectable group of novel regulatory genes and uses thereof
US8163896B1 (en) 2002-11-14 2012-04-24 Rosetta Genomics Ltd. Bioinformatically detectable group of novel regulatory genes and uses thereof
US20090227780A1 (en) * 2002-11-14 2009-09-10 Dharmacon, Inc. siRNA targeting connexin 43
US9879266B2 (en) 2002-11-14 2018-01-30 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US10920226B2 (en) * 2002-11-14 2021-02-16 Thermo Fisher Scientific Inc. siRNA targeting LDHA
US9228186B2 (en) 2002-11-14 2016-01-05 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US7635770B2 (en) * 2002-11-14 2009-12-22 Dharmacon, Inc. siRNA targeting protein kinase N-3 (PKN-3)
US8198427B1 (en) 2002-11-14 2012-06-12 Dharmacon, Inc. SiRNA targeting catenin, beta-1 (CTNNB1)
US20100113307A1 (en) * 2002-11-14 2010-05-06 Dharmacon, Inc. siRNA targeting vascular endothelial growth factor (VEGF)
WO2006006948A2 (en) 2002-11-14 2006-01-19 Dharmacon, Inc. METHODS AND COMPOSITIONS FOR SELECTING siRNA OF IMPROVED FUNCTIONALITY
US20090005548A1 (en) * 2002-11-14 2009-01-01 Dharmacon, Inc. siRNA targeting nuclear receptor interacting protein 1 (NRIP1)
US7592442B2 (en) * 2002-11-14 2009-09-22 Dharmacon, Inc. siRNA targeting ribonucleotide reductase M2 polypeptide (RRM2 or RNR-R2)
US9719094B2 (en) 2002-11-14 2017-08-01 Thermo Fisher Scientific Inc. RNAi targeting SEC61G
US7781575B2 (en) 2002-11-14 2010-08-24 Dharmacon, Inc. siRNA targeting tumor protein 53 (p53)
US20080268457A1 (en) * 2002-11-14 2008-10-30 Dharmacon, Inc. siRNA targeting forkhead box P3 (FOXP3)
US9719092B2 (en) 2002-11-14 2017-08-01 Thermo Fisher Scientific Inc. RNAi targeting CNTD2
US7951935B2 (en) 2002-11-14 2011-05-31 Dharmacon, Inc. siRNA targeting v-myc myelocytomatosis viral oncogene homolog (MYC)
US9839649B2 (en) 2002-11-14 2017-12-12 Thermo Fisher Scientific Inc. Methods and compositions for selecting siRNA of improved functionality
US7977471B2 (en) * 2002-11-14 2011-07-12 Dharmacon, Inc. siRNA targeting TNFα
PT2284266E (pt) * 2002-11-14 2013-12-17 Thermo Fisher Scient Biosciences Inc Siarn contra tp53
US7619081B2 (en) * 2002-11-14 2009-11-17 Dharmacon, Inc. siRNA targeting coatomer protein complex, subunit beta 2 (COPB2)
US9771586B2 (en) 2002-11-14 2017-09-26 Thermo Fisher Scientific Inc. RNAi targeting ZNF205
CA2506619A1 (en) 2002-11-18 2004-08-19 Thomas W. Hodge Cell lines and host nucleic acid sequences related to infectious disease
US7064337B2 (en) 2002-11-19 2006-06-20 The Regents Of The University Of California Radiation detection system for portable gamma-ray spectroscopy
DE10254214A1 (de) * 2002-11-20 2004-06-09 Beiersdorf Ag Oligoribonukleotide zur Behandlung von degenerativen Hauterscheinungen durch RNA-Interferenz
WO2004047764A2 (en) * 2002-11-22 2004-06-10 University Of Massachusetts Modulation of hiv replication by rna interference
WO2004048566A1 (ja) * 2002-11-22 2004-06-10 Bio-Think Tank Co., Ltd. Rna干渉の標的塩基配列検索方法、rna干渉を生じさせるポリヌクレオチドの塩基配列設計方法、2本鎖ポリヌクレオチドの作製方法、遺伝子の発現抑制方法、塩基配列処理装置、塩基配列処理方法をコンピュータに実行させるプログラム、記録媒体、および、塩基配列処理システム
JP4526228B2 (ja) * 2002-11-22 2010-08-18 隆 森田 RNAiによる新規治療法および治療剤
US7829694B2 (en) 2002-11-26 2010-11-09 Medtronic, Inc. Treatment of neurodegenerative disease through intracranial delivery of siRNA
US7790867B2 (en) 2002-12-05 2010-09-07 Rosetta Genomics Inc. Vaccinia virus-related nucleic acids and microRNA
US20130130231A1 (en) 2002-11-26 2013-05-23 Isaac Bentwich Bioinformatically detectable group of novel viral regulatory genes and uses thereof
US7696334B1 (en) 2002-12-05 2010-04-13 Rosetta Genomics, Ltd. Bioinformatically detectable human herpesvirus 5 regulatory gene
US7605249B2 (en) 2002-11-26 2009-10-20 Medtronic, Inc. Treatment of neurodegenerative disease through intracranial delivery of siRNA
US7618948B2 (en) 2002-11-26 2009-11-17 Medtronic, Inc. Devices, systems and methods for improving and/or cognitive function through brain delivery of siRNA
CA2506714A1 (en) * 2002-11-26 2004-06-10 University Of Massachusetts Delivery of sirnas
CN1301263C (zh) 2002-12-18 2007-02-21 北京昭衍新药研究中心 一组抗hiv感染及防治艾滋病的核苷酸序列及其应用
DK2270048T3 (en) 2002-12-24 2016-01-18 Rinat Neuroscience Corp Anti-NGF antibodies and methods for their use
US9498530B2 (en) 2002-12-24 2016-11-22 Rinat Neuroscience Corp. Methods for treating osteoarthritis pain by administering a nerve growth factor antagonist and compositions containing the same
EP1590430A4 (en) * 2003-01-03 2009-08-05 Gencia Corp SiRNA-mediated post-transcriptional gene silencing of allopecia genes
MXPA05007651A (es) 2003-01-16 2005-10-26 Univ Pennsylvania Composiciones y metodos para la inhibicion por arnsi de la molecula de adhesion intercelular-1.
US7629323B2 (en) * 2003-01-21 2009-12-08 Northwestern University Manipulation of neuronal ion channels
US20060178297A1 (en) * 2003-01-28 2006-08-10 Troy Carol M Systems and methods for silencing expression of a gene in a cell and uses thereof
US20040147027A1 (en) * 2003-01-28 2004-07-29 Troy Carol M. Complex for facilitating delivery of dsRNA into a cell and uses thereof
US7994149B2 (en) 2003-02-03 2011-08-09 Medtronic, Inc. Method for treatment of Huntington's disease through intracranial delivery of sirna
US7732591B2 (en) 2003-11-25 2010-06-08 Medtronic, Inc. Compositions, devices and methods for treatment of huntington's disease through intracranial delivery of sirna
CA2514912A1 (en) * 2003-02-05 2004-08-26 University Of Massachusetts Rnai targeting of viruses
FR2850971B1 (fr) * 2003-02-10 2006-08-11 Aventis Pharma Sa Oligonucleotide antisens inhibant l'expression de la proteine ob-rgrp et procede de detection de composes modifiant l'interaction entre la famille de la proteine ob-rgrp et le recepteur de la leptine
US20070104688A1 (en) 2003-02-13 2007-05-10 City Of Hope Small interfering RNA mediated transcriptional gene silencing in mammalian cells
US20040162235A1 (en) * 2003-02-18 2004-08-19 Trubetskoy Vladimir S. Delivery of siRNA to cells using polyampholytes
EP1594441B1 (en) 2003-02-19 2010-12-15 Rinat Neuroscience Corp. Method for treating pain by administering a nerve growth factor antagonist and an nsaid and composition containing the same
WO2004076664A2 (en) * 2003-02-21 2004-09-10 University Of South Florida Vectors for regulating gene expression
EP1611231A4 (en) * 2003-02-21 2008-08-13 Penn State Res Found RNA-INTERFERING COMPOSITIONS AND METHODS THEREOF
AU2004215081A1 (en) * 2003-02-27 2004-09-10 National Institute Of Advanced Industrial Science And Technology Induction of methylation of CpG sequence by dsRNA in mammalian cell
ATE554185T1 (de) * 2003-02-27 2012-05-15 Alnylam Pharmaceuticals Inc Verfahren und konstrukte zur bewertung von rnai- zielen und effektormolekülen
JP2006520611A (ja) * 2003-03-05 2006-09-14 セネスコ テクノロジーズ,インコーポレイティド eIF−5A1の発現を抑制するための、アンチセンス・オリゴヌクレオチド又はsiRNAの使用
WO2004078941A2 (en) * 2003-03-06 2004-09-16 Oligo Engine, Inc. Modulation of gene expression using dna-rna hybrids
ATE479752T1 (de) 2003-03-07 2010-09-15 Alnylam Pharmaceuticals Inc Therapeutische zusammensetzungen
EP1605961A4 (en) * 2003-03-12 2009-11-11 Vasgene Therapeutics Inc POLYPEPTIDE COMPOUNDS FOR INHIBITING ANGIOGENESIS AND TUMOR GROWTH
DE602004023279D1 (de) 2003-03-21 2009-11-05 Santaris Pharma As Analoga kurzer interferierender rna (sirna)
US20040198640A1 (en) * 2003-04-02 2004-10-07 Dharmacon, Inc. Stabilized polynucleotides for use in RNA interference
JP4605799B2 (ja) * 2003-04-02 2011-01-05 ダーマコン, インコーポレイテッド Rna干渉において使用するための修飾ポリヌクレオチド
ATE536408T1 (de) * 2003-04-02 2011-12-15 Dharmacon Inc Modifizierte polynukleotide zur verwendung bei rna-interferenz
US20050013855A1 (en) 2003-04-09 2005-01-20 Biodelivery Sciences International, Inc. Cochleate compositions directed against expression of proteins
WO2004091515A2 (en) 2003-04-09 2004-10-28 Alnylam Pharmaceuticals, Inc. iRNA CONJUGATES
WO2004092383A2 (en) * 2003-04-15 2004-10-28 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF SEVERE ACUTE RESPIRATORY SYNDROME (SARS) VIRUS GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
EP2660322A3 (en) 2003-04-17 2013-11-13 Alnylam Pharmaceuticals Inc. Modified iRNA agents
US8796436B2 (en) 2003-04-17 2014-08-05 Alnylam Pharmaceuticals, Inc. Modified iRNA agents
US7723509B2 (en) 2003-04-17 2010-05-25 Alnylam Pharmaceuticals IRNA agents with biocleavable tethers
US8017762B2 (en) 2003-04-17 2011-09-13 Alnylam Pharmaceuticals, Inc. Modified iRNA agents
US7851615B2 (en) 2003-04-17 2010-12-14 Alnylam Pharmaceuticals, Inc. Lipophilic conjugated iRNA agents
AU2004232964B2 (en) 2003-04-17 2011-09-22 Alnylam Pharmaceuticals, Inc. Protected monomers
AU2004233043A1 (en) * 2003-04-18 2004-11-04 The Trustees Of The University Of Pennsylvania Compositions and methods for siRNA inhibition of angiopoietin 1 and 2 and their receptor Tie2
US20070010468A1 (en) * 2003-04-23 2007-01-11 Georgetown University Methods and compositions for the inhibition of stat5 in prostate cancer cells
US9701725B2 (en) * 2003-05-05 2017-07-11 The Johns Hopkins University Anti-cancer DNA vaccine employing plasmids encoding signal sequence, mutant oncoprotein antigen, and heat shock protein
EP1623032A2 (en) 2003-05-09 2006-02-08 University of Pittsburgh of the Commonwealth System of Higher Education Small interfering rna libraries and methods of synthesis and use
WO2004101756A2 (en) 2003-05-09 2004-11-25 Diadexus, Inc. Ovr110 antibody compositions and methods of use
US7964714B2 (en) * 2003-05-12 2011-06-21 Potomac Pharmaceuticals Inc. Gene expression suppression agents
US20050148531A1 (en) * 2003-05-15 2005-07-07 Todd Hauser Modulation of gene expression using DNA-DNA hybrids
WO2005018534A2 (en) * 2003-05-16 2005-03-03 Rosetta Inpharmatics, Llc Methods and compositions for rna interference
WO2004100990A1 (ja) * 2003-05-19 2004-11-25 Genecare Research Institute Co., Ltd. 癌細胞に対するアポトーシス誘導剤
AU2005212433B2 (en) * 2003-05-23 2010-12-16 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using multifunctional short interfering nucleic acid (multifunctional sINA)
WO2004105774A1 (ja) * 2003-05-30 2004-12-09 Nippon Shinyaku Co., Ltd. オリゴ核酸担持複合体、当該複合体を含有する医薬組成物
KR20060013426A (ko) 2003-05-30 2006-02-09 니뽄 신야쿠 가부시키가이샤 Bcl-2의 발현을 억제하는 올리고 이중쇄 RNA와그것을 함유하는 의약 조성물
PT1633767T (pt) * 2003-06-02 2019-02-27 Univ Massachusetts Métodos e composições para controlar a eficácia do silenciamento de arn
US7750144B2 (en) 2003-06-02 2010-07-06 University Of Massachusetts Methods and compositions for enhancing the efficacy and specificity of RNA silencing
ES2864206T3 (es) * 2003-06-02 2021-10-13 Univ Massachusetts Métodos y composiciones para mejorar la eficacia y la especificidad del ARNi
US20050019918A1 (en) * 2003-06-03 2005-01-27 Hidetoshi Sumimoto Treatment of cancer by inhibiting BRAF expression
BRPI0410886A (pt) * 2003-06-03 2006-07-04 Isis Pharmaceuticals Inc composto de filamento duplo, composição farmacêutica, sal farmaceuticamente aceitável, métodos de modificação do ácido nucleico que codifica a survivina humana, de inibição da expressão da suvivina em células ou tecidos, e de tratamento de uma condição associada com a expressão ou superexpressão da suvivina, e, oligonucleotìdeo de rnai de filamento único
US7595306B2 (en) * 2003-06-09 2009-09-29 Alnylam Pharmaceuticals Inc Method of treating neurodegenerative disease
US8575327B2 (en) 2003-06-12 2013-11-05 Alnylam Pharmaceuticals, Inc. Conserved HBV and HCV sequences useful for gene silencing
DK2336317T3 (da) * 2003-06-13 2019-12-16 Alnylam Europe Ag Dobbeltstrenget ribonukleinsyre med forøget effektivitet i en organisme
EP1486564A1 (de) * 2003-06-13 2004-12-15 Ribopharma AG SiRNA mit erhöhter Stabilität in Serum
WO2005044976A2 (en) * 2003-06-20 2005-05-19 Isis Pharmaceuticals, Inc. Oligomeric compounds for use in gene modulation
US7790691B2 (en) * 2003-06-20 2010-09-07 Isis Pharmaceuticals, Inc. Double stranded compositions comprising a 3′-endo modified strand for use in gene modulation
US7173015B2 (en) * 2003-07-03 2007-02-06 The Trustees Of The University Of Pennsylvania Inhibition of Syk kinase expression
EP1649019A2 (en) * 2003-07-15 2006-04-26 California Institute Of Technology Improved inhibitor nucleic acids
US20050256071A1 (en) * 2003-07-15 2005-11-17 California Institute Of Technology Inhibitor nucleic acids
CA2532228C (en) * 2003-07-16 2017-02-14 Protiva Biotherapeutics, Inc. Lipid encapsulated interfering rna
WO2005010188A2 (en) * 2003-07-21 2005-02-03 Whitehead Institute For Biomedical Research Rnas able to modulate chromatin silencing
EP1648914A4 (en) 2003-07-31 2009-12-16 Regulus Therapeutics Inc OLIGOMERIC COMPOUNDS AND COMPOSITIONS USEFUL FOR MODULATING SMALL NON-CODING RNA
US20060229266A1 (en) * 2003-08-13 2006-10-12 Kumar Nalin M Silencing of tgf-beta receptor type II expression by sirna
US7888497B2 (en) 2003-08-13 2011-02-15 Rosetta Genomics Ltd. Bioinformatically detectable group of novel regulatory oligonucleotides and uses thereof
US7825235B2 (en) * 2003-08-18 2010-11-02 Isis Pharmaceuticals, Inc. Modulation of diacylglycerol acyltransferase 2 expression
WO2005035759A2 (en) * 2003-08-20 2005-04-21 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF HYPOXIA INDUCIBLE FACTOR 1 (HIF1) GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
US20050136437A1 (en) * 2003-08-25 2005-06-23 Nastech Pharmaceutical Company Inc. Nanoparticles for delivery of nucleic acids and stable double-stranded RNA
EP1660657A1 (en) * 2003-08-28 2006-05-31 Novartis AG Interfering rna duplex having blunt-ends and 3'-modifications
US8501705B2 (en) * 2003-09-11 2013-08-06 The Board Of Regents Of The University Of Texas System Methods and materials for treating autoimmune and/or complement mediated diseases and conditions
EP2821085B1 (en) * 2003-09-12 2020-04-29 University of Massachusetts Rna interference for the treatment of gain-of-function disorders
US8680063B2 (en) * 2003-09-12 2014-03-25 University Of Massachusetts RNA interference for the treatment of gain-of-function disorders
KR100750788B1 (ko) 2003-09-18 2007-08-20 아이시스 파마수티컬즈 인코포레이티드 eIF4E 발현의 조정
JP2007505634A (ja) * 2003-09-22 2007-03-15 ロゼッタ インファーマティクス エルエルシー Rna干渉を用いる合成致死スクリーニング
WO2005033310A1 (de) * 2003-10-01 2005-04-14 Grünenthal GmbH Pim-1-spezifische dsrna-verbindungen
EP1670425A4 (en) * 2003-10-07 2008-04-16 Quark Pharmaceuticals Inc BONE MORPHOGENETIC PROTEIN (BMP) 2A AND ITS APPLICATIONS
WO2005035769A2 (en) 2003-10-09 2005-04-21 E. I. Du Pont De Nemours And Company Gene silencing by using micro-rna molecules
WO2005045032A2 (en) * 2003-10-20 2005-05-19 Sima Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF EARLY GROWTH RESPONSE GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
WO2005045038A2 (en) * 2003-10-23 2005-05-19 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF GPRA AND AAA1 GENE EXPRESSION USING SHORT NUCLEIC ACID (siNA)
EP1675949A2 (en) * 2003-10-23 2006-07-05 Sirna Therapeutics, Inc. RNA INTERFERENCE MEDIATED INHIBITION OF NOGO AND NOGO RECEPTOR GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
AU2004286261B2 (en) 2003-10-27 2010-06-24 Merck Sharp & Dohme Llc Method of designing siRNAs for gene silencing
US8227434B1 (en) 2003-11-04 2012-07-24 H. Lee Moffitt Cancer Center & Research Institute, Inc. Materials and methods for treating oncological disorders
WO2005047504A1 (en) * 2003-11-07 2005-05-26 The Board Of Trustees Of The University Of Illinois Induction of cellular senescence by cdk4 disruption for tumor suppression and regression
EP1697515A4 (en) * 2003-11-12 2008-02-06 Austin Research Inst CONJUGATE DNA-EXCIPIENT
US7807646B1 (en) * 2003-11-20 2010-10-05 University Of South Florida SHIP-deficiency to increase megakaryocyte progenitor production
JP2005168485A (ja) * 2003-11-20 2005-06-30 Tsutomu Suzuki siRNAの設計方法
US7763592B1 (en) 2003-11-20 2010-07-27 University Of South Florida SHIP-deficiency to increase megakaryocyte progenitor production
US20050208658A1 (en) * 2003-11-21 2005-09-22 The University Of Maryland RNA interference mediated inhibition of 11beta hydroxysteriod dehydrogenase-1 (11beta HSD-1) gene expression
US20100145038A1 (en) * 2003-11-24 2010-06-10 Merck & Co., Inc. RNA INTERFERENCE MEDIATED INHIBITION OF GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
AU2004294567A1 (en) * 2003-11-26 2005-06-16 University Of Massachusetts Sequence-specific inhibition of small RNA function
US20080171051A1 (en) * 2003-11-26 2008-07-17 Patrick Gerard Johnston Cancer Treatment
WO2005056021A1 (en) 2003-12-04 2005-06-23 University Of South Florida Polynucleotides for reducing respiratory syncytial virus gene expression
US20050234000A1 (en) * 2003-12-12 2005-10-20 Mitchell Gordon S SiRNA delivery into mammalian nerve cells
JPWO2005068630A1 (ja) * 2003-12-16 2007-07-26 独立行政法人産業技術総合研究所 干渉用二重鎖rna
US20060134787A1 (en) * 2004-12-22 2006-06-22 University Of Massachusetts Methods and compositions for enhancing the efficacy and specificity of single and double blunt-ended siRNA
CA2551100A1 (en) * 2003-12-23 2005-07-14 The Trustees Of The University Of Pennsylvania Compositions and methods for combined therapy of disease
JP4767019B2 (ja) * 2004-01-16 2011-09-07 武田薬品工業株式会社 動脈硬化の予防・治療用医薬
EP1713912B1 (en) * 2004-01-30 2013-09-18 Santaris Pharma A/S Modified short interfering rna (modified sirna)
DE602005025347D1 (de) * 2004-01-30 2011-01-27 Quark Pharmaceuticals Inc Oligoribonukleotide und verfahren zu deren anwendung bei der behandlung von fibrotischen leiden und anderen krankheiten
ATE447024T1 (de) * 2004-02-06 2009-11-15 Dharmacon Inc Stabilisierte rnas als transfektionskontrollen und silencing-reagentien
WO2005078848A2 (en) 2004-02-11 2005-08-25 University Of Tennessee Research Foundation Inhibition of tumor growth and invasion by anti-matrix metalloproteinase dnazymes
US20060069050A1 (en) * 2004-02-17 2006-03-30 University Of Massachusetts Methods and compositions for mediating gene silencing
WO2005079532A2 (en) * 2004-02-17 2005-09-01 University Of Massachusetts Methods and compositions for enhancing risc activity in vitro and in vivo
JP2007523214A (ja) * 2004-02-23 2007-08-16 ジェンザイム コーポレイション デスレセプターリガンド誘導アポトーシスのmuc1アンタゴニスト増強方法
EP1566202A1 (en) * 2004-02-23 2005-08-24 Sahltech I Göteborg AB Use of resistin antagonists in the treatment of rheumatoid arthritis
WO2005092924A2 (en) 2004-02-24 2005-10-06 The Government Of The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services, Centers For Disease Control And Prevention Rab9a, rab11a, and modulators thereof related to infectious disease
US7691823B2 (en) * 2004-03-05 2010-04-06 University Of Massachusetts RIP140 regulation of glucose transport
US8569474B2 (en) 2004-03-09 2013-10-29 Isis Pharmaceuticals, Inc. Double stranded constructs comprising one or more short strands hybridized to a longer strand
US20050202075A1 (en) * 2004-03-12 2005-09-15 Pardridge William M. Delivery of genes encoding short hairpin RNA using receptor-specific nanocontainers
JP4937899B2 (ja) 2004-03-12 2012-05-23 アルナイラム ファーマシューティカルズ, インコーポレイテッド VEGFを標的とするiRNA物質
US20070265220A1 (en) * 2004-03-15 2007-11-15 City Of Hope Methods and compositions for the specific inhibition of gene expression by double-stranded RNA
EP1742958B1 (en) 2004-03-15 2017-05-17 City of Hope Methods and compositions for the specific inhibition of gene expression by double-stranded rna
US20050208090A1 (en) * 2004-03-18 2005-09-22 Medtronic, Inc. Methods and systems for treatment of neurological diseases of the central nervous system
US7851452B2 (en) * 2004-03-22 2010-12-14 The Trustees Of The University Of Pennsylvania Methods of use of bcl-6-derived nucleotides to induce apoptosis
US20050272682A1 (en) * 2004-03-22 2005-12-08 Evers Bernard M SiRNA targeting PI3K signal transduction pathway and siRNA-based therapy
US7872117B2 (en) * 2004-03-26 2011-01-18 Van Andel Research Institute c-met siRNA adenovirus vectors inhibit cancer cell growth, invasion and tumorigenicity
EP1730280B1 (en) * 2004-03-26 2018-10-24 Curis, Inc. Rna interference modulators of hedgehog signaling and uses thereof
JP2005312428A (ja) * 2004-03-31 2005-11-10 Keio Gijuku Skp−2発現抑制を利用した癌の治療
WO2005095647A1 (ja) * 2004-03-31 2005-10-13 Takara Bio Inc. siRNAのスクリーニング方法
KR101147147B1 (ko) * 2004-04-01 2012-05-25 머크 샤프 앤드 돔 코포레이션 Rna 간섭의 오프 타겟 효과 감소를 위한 변형된폴리뉴클레오타이드
US20050267300A1 (en) 2004-04-05 2005-12-01 Muthiah Manoharan Processes and reagents for oligonucleotide synthesis and purification
EP2481802B1 (en) 2004-04-09 2015-06-10 Genecare Research Institute Co., Ltd Cancer cell-specific apoptosis-inducing agents that target chromosome stabilization-associated genes
US20060078902A1 (en) * 2004-04-15 2006-04-13 Michaeline Bunting Method and compositions for RNA interference
US20060014289A1 (en) * 2004-04-20 2006-01-19 Nastech Pharmaceutical Company Inc. Methods and compositions for enhancing delivery of double-stranded RNA or a double-stranded hybrid nucleic acid to regulate gene expression in mammalian cells
AU2005238034A1 (en) 2004-04-23 2005-11-10 The Trustees Of Columbia University In The City Of New York Inhibition of hairless protein mRNA
US7626014B2 (en) 2004-04-27 2009-12-01 Alnylam Pharmaceuticals Single-stranded and double-stranded oligonucleotides comprising a 2-arylpropyl moiety
CA2564868C (en) 2004-04-28 2013-11-26 Molecules For Health, Inc. Methods for treating or preventing restenosis and other vascular proliferative disorders
EP3034510A1 (en) 2004-04-30 2016-06-22 Alnylam Pharmaceuticals Inc. Oligonucleotides comprising a c5-modified pyrimidine
US20060040882A1 (en) * 2004-05-04 2006-02-23 Lishan Chen Compostions and methods for enhancing delivery of nucleic acids into cells and for modifying expression of target genes in cells
US20110117088A1 (en) * 2004-05-12 2011-05-19 Simon Michael R Composition and method for introduction of rna interference sequences into targeted cells and tissues
US7605250B2 (en) * 2004-05-12 2009-10-20 Dharmacon, Inc. siRNA targeting cAMP-specific phosphodiesterase 4D
US20060030003A1 (en) * 2004-05-12 2006-02-09 Simon Michael R Composition and method for introduction of RNA interference sequences into targeted cells and tissues
US20050260214A1 (en) * 2004-05-12 2005-11-24 Simon Michael R Composition and method for introduction of RNA interference sequences into targeted cells and tissues
WO2005110464A2 (en) * 2004-05-14 2005-11-24 Oregon Health & Science University Irx5 inhibition as treatment for hyperproliferative disorders
CA2566519C (en) 2004-05-14 2020-04-21 Rosetta Genomics Ltd. Micrornas and uses thereof
US7687616B1 (en) 2004-05-14 2010-03-30 Rosetta Genomics Ltd Small molecules modulating activity of micro RNA oligonucleotides and micro RNA targets and uses thereof
US10508277B2 (en) 2004-05-24 2019-12-17 Sirna Therapeutics, Inc. Chemically modified multifunctional short interfering nucleic acid molecules that mediate RNA interference
EP1773303A2 (en) * 2004-05-25 2007-04-18 Chimeracore, Inc. Self-assembling nanoparticle drug delivery system
US7795419B2 (en) 2004-05-26 2010-09-14 Rosetta Genomics Ltd. Viral and viral associated miRNAs and uses thereof
EP2471924A1 (en) 2004-05-28 2012-07-04 Asuragen, INC. Methods and compositions involving microRNA
US8394947B2 (en) 2004-06-03 2013-03-12 Isis Pharmaceuticals, Inc. Positionally modified siRNA constructs
EP1602926A1 (en) * 2004-06-04 2005-12-07 University of Geneva Novel means and methods for the treatment of hearing loss and phantom hearing
JP4796062B2 (ja) * 2004-06-07 2011-10-19 プロチバ バイオセラピューティクス インコーポレイティッド 脂質封入干渉rna
WO2005120152A2 (en) * 2004-06-07 2005-12-22 Protiva Biotherapeutics, Inc. Cationic lipids and methods of use
US20060008907A1 (en) * 2004-06-09 2006-01-12 The Curators Of The University Of Missouri Control of gene expression via light activated RNA interference
US20090215860A1 (en) * 2004-06-17 2009-08-27 The Regents Of The University Of California Compositions and methods for regulating gene transcription
US20060051815A1 (en) * 2004-06-25 2006-03-09 The J. David Gladstone Institutes Methods of treating smooth muscle cell disorders
WO2006088490A2 (en) 2004-06-30 2006-08-24 Alnylam Pharmaceuticals, Inc. Oligonucleotides comprising a non-phosphate backbone linkage
CA2572439A1 (en) * 2004-07-02 2006-01-12 Protiva Biotherapeutics, Inc. Immunostimulatory sirna molecules and uses therefor
EP1765378B1 (en) 2004-07-12 2014-04-16 Medical Research Fund of Tel Aviv Sourasky Medical Center Agent capable of downregulating an msf-a-dependent hif-1a and use thereof in cancer treatment
CA2574572A1 (en) * 2004-07-19 2006-10-26 Baylor College Of Medicine Modulation of cytokine signaling regulators and applications for immunotherapy
WO2006020231A2 (en) * 2004-07-21 2006-02-23 Medtronic, Inc. Medical devices and methods for reducing localized fibrosis
WO2006093526A2 (en) 2004-07-21 2006-09-08 Alnylam Pharmaceuticals, Inc. Oligonucleotides comprising a modified or non-natural nucleobase
WO2006020230A2 (en) * 2004-07-21 2006-02-23 Medtronic, Inc. METHODS FOR REDUCING OR PREVENTING LOCALIZED FIBROSIS USING SiRNA
US20100132058A1 (en) 2004-07-23 2010-05-27 Diatchenko Luda B Methods and materials for determining pain sensitivity and predicting and treating related disorders
EP1913011B1 (en) 2004-08-04 2016-11-02 Alnylam Pharmaceuticals Inc. Oligonucleotides comprising a ligand tethered to a modified or non-natural nucleobase
EP1791567B1 (en) 2004-08-10 2015-07-29 Alnylam Pharmaceuticals Inc. Chemically modified oligonucleotides
WO2006020557A2 (en) * 2004-08-10 2006-02-23 Immusol, Inc. Methods of using or identifying agents that inhibit cancer growth
WO2006110161A2 (en) * 2004-08-13 2006-10-19 University Of Delaware Method for identification and quantification of short or small rna molecules
DK2319925T3 (da) 2004-08-16 2018-11-05 Quark Pharmaceuticals Inc Terapeutiske anvendelser af RTP801-hæmmere
US20070021366A1 (en) * 2004-11-19 2007-01-25 Srivastava Satish K Structural-based inhibitors of the glutathione binding site in aldose reductase, methods of screening therefor and methods of use
CN105251024A (zh) * 2004-08-23 2016-01-20 西伦蒂斯私人股份公司 眼病的治疗
WO2006024880A2 (en) * 2004-08-31 2006-03-09 Sylentis S.A.U. Methods and compositions to inhibit p2x7 receptor expression
WO2006026738A2 (en) 2004-08-31 2006-03-09 Qiagen North American Holdings, Inc. Methods and compositions for rna amplification and detection using an rna-dependent rna-polymerase
US7884086B2 (en) * 2004-09-08 2011-02-08 Isis Pharmaceuticals, Inc. Conjugates for use in hepatocyte free uptake assays
EP1793835A4 (en) 2004-09-10 2010-12-01 Somagenics Inc SMALL INTERFERING RNA EFFECTIVELY INHIBITING VIRAL GENE EXPRESSION AND METHODS OF USE THEREOF
WO2006033965A2 (en) * 2004-09-16 2006-03-30 The Trustees Of The University Of Pennsylvania Nadph oxidase inhibition pharmacotherapies for obstructive sleep apnea syndrome and its associated morbidities
FI20041204A0 (fi) 2004-09-16 2004-09-16 Riikka Lund Menetelmät immuunivälitteisiin sairauksiin liittyvien uusien kohdegeenien hyödyntämiseksi
CN102628044A (zh) 2004-09-24 2012-08-08 阿尔尼拉姆医药品有限公司 APOB的RNAi调节及其用途
BRPI0516177B8 (pt) 2004-09-28 2021-05-25 Quark Biotech Inc oligorribonucleotídeos, uso dos mesmos, uso de um composto sirna eficaz para inibir 0 p53 humano e composição farmacêutica que os contém, bem como método para diminuição da expressão do gene p53
WO2006039343A2 (en) * 2004-09-30 2006-04-13 Centocor, Inc. Emmprin antagonists and uses thereof
EP2336333A1 (en) 2004-10-21 2011-06-22 Venganza Inc. Methods and materials for conferring resistance to pests and pathogens of plants
US20060089324A1 (en) * 2004-10-22 2006-04-27 Sailen Barik RNAi modulation of RSV, PIV and other respiratory viruses and uses thereof
US20060110440A1 (en) * 2004-10-22 2006-05-25 Kiminobu Sugaya Method and system for biasing cellular development
US7790878B2 (en) * 2004-10-22 2010-09-07 Alnylam Pharmaceuticals, Inc. RNAi modulation of RSV, PIV and other respiratory viruses and uses thereof
CA2584921A1 (en) * 2004-10-22 2006-04-27 Neuregenix Limited Neuron regeneration
CN101103112B (zh) * 2004-10-27 2011-11-16 先灵公司 抑制Nav1.8的短干扰核酸组合物和方法
SG156690A1 (en) 2004-10-27 2009-11-26 Univ Vanderbilt Mammalian genes involved in infection
WO2006050002A2 (en) * 2004-10-28 2006-05-11 Idexx Laboratories, Inc. Compositions for controlled delivery of pharmaceutically active compounds
US20060094676A1 (en) * 2004-10-29 2006-05-04 Ronit Lahav Compositions and methods for treating cancer using compositions comprising an inhibitor of endothelin receptor activity
WO2007001448A2 (en) 2004-11-04 2007-01-04 Massachusetts Institute Of Technology Coated controlled release polymer particles as efficient oral delivery vehicles for biopharmaceuticals
ES2534303T3 (es) 2004-11-12 2015-04-21 Asuragen, Inc. Procedimientos y composiciones que implican miARN y moléculas inhibidoras de miARN
JP2008520583A (ja) * 2004-11-15 2008-06-19 マウント シナイ スクール オブ メディスン オブ ニューヨーク ユニバーシティー Wnt自己分泌シグナル伝達を改変するための組成物および方法
US20060105052A1 (en) * 2004-11-15 2006-05-18 Acar Havva Y Cationic nanoparticle having an inorganic core
CA2587411A1 (en) * 2004-11-17 2006-05-26 Protiva Biotherapeutics, Inc. Sirna silencing of apolipoprotein b
PT2189469E (pt) * 2004-11-18 2016-01-22 Univ Illinois Construções de arnsi multicistrónicas para inibir tumores
US8314073B2 (en) * 2004-11-19 2012-11-20 Genecare Research Institute Co., Ltd. Cancer-cell-specific cell proliferation inhibitors
US7923207B2 (en) 2004-11-22 2011-04-12 Dharmacon, Inc. Apparatus and system having dry gene silencing pools
US7935811B2 (en) 2004-11-22 2011-05-03 Dharmacon, Inc. Apparatus and system having dry gene silencing compositions
US20060166234A1 (en) * 2004-11-22 2006-07-27 Barbara Robertson Apparatus and system having dry control gene silencing compositions
AU2005323303A1 (en) * 2004-11-24 2006-07-13 Alnylam Pharmaceuticals, Inc. RNAi modulation of the Bcr-Abl fusion gene and uses thereof
JP2008521909A (ja) * 2004-12-02 2008-06-26 ビー−ブリッジ インターナショナル,インコーポレーテッド 短鎖干渉rna、アンチセンスポリヌクレオチド、および他のハイブリッド形成化ポリヌクレオチドの設計方法
WO2006060779A2 (en) * 2004-12-03 2006-06-08 Case Western Reserve University Novel methods, compositions and devices for inducing neovascularization
WO2006066158A2 (en) * 2004-12-14 2006-06-22 Alnylam Pharmaceuticals, Inc. Rnai modulation of mll-af4 and uses thereof
EP2270136A1 (en) 2004-12-17 2011-01-05 Beth Israel Deaconess Medical Center Compositions for bacterial mediated gene silencing and methods of using same
GB0427916D0 (en) * 2004-12-21 2005-01-19 Astrazeneca Ab Method
US20060142228A1 (en) * 2004-12-23 2006-06-29 Ambion, Inc. Methods and compositions concerning siRNA's as mediators of RNA interference
TWI386225B (zh) 2004-12-23 2013-02-21 Alcon Inc 用於治療眼睛病症的結締組織生長因子(CTGF)RNA干擾(RNAi)抑制技術
KR101366482B1 (ko) 2004-12-27 2014-02-21 사일런스 테라퓨틱스 아게 코팅된 지질 복합체 및 이들의 용도
US20090005332A1 (en) * 2004-12-30 2009-01-01 Hauser Todd M Compositions and Methods for Modulating Gene Expression Using Self-Protected Oligonucleotides
WO2006073970A2 (en) * 2005-01-06 2006-07-13 The Johns Hopkins University Rna interference that blocks expression of pro-apoptotic proteins potentiates immunity induced by dna and transfected dendritic cell vaccines
US7507809B2 (en) * 2005-01-07 2009-03-24 Alnylam Pharmaceuticals, Inc. RNAi modulation of RSV and therapeutic uses thereof
EP1841793B1 (en) 2005-01-07 2010-03-31 Diadexus, Inc. Ovr110 antibody compositions and methods of use
EP1841464B1 (en) * 2005-01-24 2012-06-27 Alnylam Pharmaceuticals Inc. Rnai modulation of the nogo-l or nogo-r gene and uses thereof
WO2006081323A2 (en) * 2005-01-26 2006-08-03 The Johns Hopkins University Anti-cancer dna vaccine employing plasmids encoding mutant oncoprotein antigen and calreticulin
TW200639252A (en) * 2005-02-01 2006-11-16 Alcon Inc RNAi-mediated inhibition of ocular hypertension targets
KR20130100207A (ko) 2005-02-14 2013-09-09 유니버시티 오브 아이오와 리써치 파운데이션 연령­관련 황반 변성의 치료 및 진단용 방법 및 시약
EP1848805A2 (en) * 2005-02-14 2007-10-31 HVC Strategic Research Institute, Inc Pharmaceutical agents for preventing metastasis of cancer
US8859749B2 (en) 2005-03-08 2014-10-14 Qiagen Gmbh Modified short interfering RNA
MX2007010608A (es) 2005-03-11 2007-10-19 Alcon Inc Inhibicion mediada por i-arn de proteina i relacionada con frizzled para tratamiento de glaucoma.
US8999943B2 (en) * 2005-03-14 2015-04-07 Board Of Regents, The University Of Texas System Antigene oligomers inhibit transcription
GB0505081D0 (en) * 2005-03-14 2005-04-20 Genomica Sau Downregulation of interleukin-12 expression by means of rnai technology
JP4585342B2 (ja) * 2005-03-18 2010-11-24 株式会社資生堂 不全角化を抑制する物質のスクリーニング方法、同方法によりスクリーニングされた物質及び不全角化を抑制する方法
EP1877556B1 (en) * 2005-03-25 2011-09-14 Medtronic, Inc. Use of anti-tnf or anti-il1 rnai to suppress pro- inflammatory cytokine actions locally to treat pain
ATE541928T1 (de) * 2005-03-31 2012-02-15 Calando Pharmaceuticals Inc Inhibitoren der untereinheit 2 der ribonukleotidreduktase und ihre verwendungen
WO2006113743A2 (en) * 2005-04-18 2006-10-26 Massachusetts Institute Of Technology Compositions and methods for rna interference with sialidase expression and uses thereof
US20060257912A1 (en) 2005-05-06 2006-11-16 Medtronic, Inc. Methods and sequences to suppress primate huntington gene expression
US7902352B2 (en) 2005-05-06 2011-03-08 Medtronic, Inc. Isolated nucleic acid duplex for reducing huntington gene expression
JP5329949B2 (ja) * 2005-05-31 2013-10-30 エコーレ ポリテクニーク フェデラーレ デ ローザンヌ 遺伝子に基づいた薬物の細胞質送達のためのトリブロックコポリマー
WO2006130560A2 (en) * 2005-05-31 2006-12-07 The Trustees Of The University Of Pennsylvania Manipulation of pten in t cells as a strategy to modulate immune responses
AU2006252456A1 (en) * 2005-06-01 2006-12-07 Duke University Method of inhibiting intimal hyperplasia
US20100266574A1 (en) * 2005-06-10 2010-10-21 Orna Mor Oligoribonucleotides and Methods of Use Thereof for Treatment of Fibrotic Conditions and Other Diseases
US8252756B2 (en) 2005-06-14 2012-08-28 Northwestern University Nucleic acid functionalized nanoparticles for therapeutic applications
US7838503B2 (en) * 2005-06-15 2010-11-23 Children's Medical Center Corporation Methods for extending the replicative lifespan of cells
FI20050640A0 (fi) * 2005-06-16 2005-06-16 Faron Pharmaceuticals Oy Yhdisteitä amiinioksidaasista riippuvien sairauksien tai häiriöiden hoitoon tai estoon
US7868159B2 (en) * 2005-06-23 2011-01-11 Baylor College Of Medicine Modulation of negative immune regulators and applications for immunotherapy
JP5116107B2 (ja) * 2005-06-27 2013-01-09 アルナイラム ファーマシューティカルズ, インコーポレイテッド HIF−1のRNAi調節及びその治療的利用
US9133517B2 (en) 2005-06-28 2015-09-15 Medtronics, Inc. Methods and sequences to preferentially suppress expression of mutated huntingtin
JP5383186B2 (ja) * 2005-07-07 2014-01-08 イッサム リサーチ ディベロプメント カンパニー オブ ザ ヘブリュー ユニバーシティ オブ エルサレム リミテッド H19をダウンレギュレートする核酸薬剤、及びそれを使用する方法
US8703769B2 (en) 2005-07-15 2014-04-22 The University Of North Carolina At Chapel Hill Use of EGFR inhibitors to prevent or treat obesity
DE112006002611A5 (de) * 2005-07-25 2008-07-03 Technische Universität Dresden Rna-abhängige rna-polymerase, verfahren und kits zur amplifikation und / oder markierung von rna
WO2007014370A2 (en) * 2005-07-28 2007-02-01 University Of Delaware Small regulatory rnas and methods of use
US7919583B2 (en) 2005-08-08 2011-04-05 Discovery Genomics, Inc. Integration-site directed vector systems
US20070213257A1 (en) * 2005-08-12 2007-09-13 Nastech Pharmaceutical Company Inc. Compositions and methods for complexes of nucleic acids and peptides
EP1937066A4 (en) * 2005-08-18 2008-12-24 Alnylam Pharmaceuticals Inc METHOD AND COMPOSITIONS FOR THE TREATMENT OF NERVOUS DISEASES
US20070054873A1 (en) * 2005-08-26 2007-03-08 Protiva Biotherapeutics, Inc. Glucocorticoid modulation of nucleic acid-mediated immune stimulation
WO2007028065A2 (en) * 2005-08-30 2007-03-08 Isis Pharmaceuticals, Inc. Chimeric oligomeric compounds for modulation of splicing
US20090018097A1 (en) * 2005-09-02 2009-01-15 Mdrna, Inc Modification of double-stranded ribonucleic acid molecules
WO2007033058A2 (en) * 2005-09-13 2007-03-22 Trustees Of Dartmouth College Compositions and methods for regulating rna translation via cd154 ca-dinucleotide repeat
CN101268194A (zh) 2005-09-20 2008-09-17 巴斯福植物科学有限公司 使用ta-siRNA调控基因表达的方法
FR2890859B1 (fr) * 2005-09-21 2012-12-21 Oreal Oligonucleotide d'arn double brin inhibant l'expression de la tyrosinase
US8933043B2 (en) * 2005-09-30 2015-01-13 St. Jude Children's Research Hospital Methods for regulation of p53 translation and function
US8168584B2 (en) 2005-10-08 2012-05-01 Potentia Pharmaceuticals, Inc. Methods of treating age-related macular degeneration by compstatin and analogs thereof
US8080534B2 (en) 2005-10-14 2011-12-20 Phigenix, Inc Targeting PAX2 for the treatment of breast cancer
GB0521351D0 (en) * 2005-10-20 2005-11-30 Genomica Sau Modulation of TRPV expression levels
US7838658B2 (en) * 2005-10-20 2010-11-23 Ian Maclachlan siRNA silencing of filovirus gene expression
GB0521716D0 (en) * 2005-10-25 2005-11-30 Genomica Sau Modulation of 11beta-hydroxysteriod dehydrogenase 1 expression for the treatment of ocular diseases
US8076307B2 (en) * 2005-10-27 2011-12-13 National University Corporation NARA Institute of Science and Technology Formation/elongation of axon by inhibiting the expression or function of Singar and application to nerve regeneration
AU2006305886C1 (en) * 2005-10-28 2011-03-17 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of huntingtin gene
EP2395012B8 (en) 2005-11-02 2018-06-06 Arbutus Biopharma Corporation Modified siRNA molecules and uses thereof
CA2626584A1 (en) * 2005-11-04 2007-05-18 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of nav1.8 gene
CA2628477A1 (en) * 2005-11-07 2007-05-10 Bc Cancer Agency Inhibition of autophagy genes in cancer chemotherapy
AU2006311730B2 (en) 2005-11-09 2010-12-02 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of Factor V Leiden mutant gene
EP2641970B1 (en) 2005-11-17 2014-12-24 Board of Regents, The University of Texas System Modulation of gene expression by oligomers targeted to chromosomal DNA
US8603991B2 (en) 2005-11-18 2013-12-10 Gradalis, Inc. Individualized cancer therapy
US8916530B2 (en) 2005-11-18 2014-12-23 Gradalis, Inc. Individualized cancer therapy
US20080125384A1 (en) * 2005-11-21 2008-05-29 Shuewi Yang Simultaneous silencing and restoration of gene function
JP4901753B2 (ja) * 2005-11-24 2012-03-21 学校法人自治医科大学 プロヒビチン2(phb2)のミトコンドリア機能
US9267937B2 (en) 2005-12-15 2016-02-23 Massachusetts Institute Of Technology System for screening particles
ATE466081T1 (de) * 2005-12-22 2010-05-15 Opko Ophthalmics Llc Zusammensetzungen und verfahren zur regulierung eines komplementsystems
AR057252A1 (es) * 2005-12-27 2007-11-21 Alcon Mfg Ltd Inhibicion de rho quinasa mediada por arni para el tratamiento de trastornos oculares
US8673873B1 (en) * 2005-12-28 2014-03-18 Alcon Research, Ltd. RNAi-mediated inhibition of phosphodiesterase type 4 for treatment of cAMP-related ocular disorders
JP5713377B2 (ja) 2005-12-28 2015-05-07 ザ スクリプス リサーチ インスティテュート 薬物標的としての天然アンチセンスおよび非コードrna転写物
EP1973574B1 (en) * 2005-12-30 2014-04-02 Institut Gustave Roussy Use of inhibitors of scinderin and/or of ephrin-a1 for treating tumors
US20090060921A1 (en) * 2006-01-17 2009-03-05 Biolex Therapeutics, Inc. Glycan-optimized anti-cd20 antibodies
WO2007084922A2 (en) 2006-01-17 2007-07-26 Biolex Therapeutics, Inc. Compositions and methods for humanization and optimization of n-glycans in plants
DOP2007000015A (es) 2006-01-20 2007-08-31 Quark Biotech Inc Usos terapéuticos de inhibidores de rtp801
US7825099B2 (en) 2006-01-20 2010-11-02 Quark Pharmaceuticals, Inc. Treatment or prevention of oto-pathologies by inhibition of pro-apoptotic genes
US20120208824A1 (en) 2006-01-20 2012-08-16 Cell Signaling Technology, Inc. ROS Kinase in Lung Cancer
ES2539830T3 (es) 2006-01-20 2015-07-06 Cell Signaling Technology, Inc. ROS quinasa mutante y de translocación en el carcinoma pulmonar no microcítico humano
US20070259827A1 (en) * 2006-01-25 2007-11-08 University Of Massachusetts Compositions and methods for enhancing discriminatory RNA interference
JP2009524419A (ja) * 2006-01-27 2009-07-02 サンタリス ファーマ アー/エス Lna修飾したホスホロチオール化オリゴヌクレオチド
US8229398B2 (en) * 2006-01-30 2012-07-24 Qualcomm Incorporated GSM authentication in a CDMA network
US8362229B2 (en) * 2006-02-08 2013-01-29 Quark Pharmaceuticals, Inc. Tandem siRNAS
US7910566B2 (en) 2006-03-09 2011-03-22 Quark Pharmaceuticals Inc. Prevention and treatment of acute renal failure and other kidney diseases by inhibition of p53 by siRNA
FI20060246A0 (fi) 2006-03-16 2006-03-16 Jukka Westermarck Uusi kasvua stimuloiva proteiini ja sen käyttö
US20100056441A1 (en) * 2006-03-17 2010-03-04 Costa Robert H Method for Inhibiting Angiogenesis
EA015563B1 (ru) 2006-03-23 2011-08-30 Сантарис Фарма А/С Короткая внутренне сегментированная интерферирующая рнк
FR2898908A1 (fr) 2006-03-24 2007-09-28 Agronomique Inst Nat Rech Procede de preparation de cellules aviaires differenciees et genes impliques dans le maintien de la pluripotence
CA2644621A1 (en) * 2006-03-24 2007-10-04 Novartis Ag Dsrna compositions and methods for treating hpv infection
WO2007115047A2 (en) * 2006-03-29 2007-10-11 Senesco Technologies, Inc. Inhibition of hiv replication and expression of p24 with eif-5a
SG170780A1 (en) 2006-03-31 2011-05-30 Alnylam Pharmaceuticals Inc Compositions and methods for inhibiting expression of eg5 gene
CA2648099C (en) 2006-03-31 2012-05-29 The Brigham And Women's Hospital, Inc System for targeted delivery of therapeutic agents
US20090226446A1 (en) * 2006-04-06 2009-09-10 Deutsches Krebsforschungszentrum Stiftung Des Offentilchen Rechts Method to Inhibit the Propagation of an Undesired Cell Population
US8524454B2 (en) 2006-04-07 2013-09-03 The Research Foundation Of State University Of New York Transcobalamin receptor polypeptides, nucleic acids, and modulators thereof, and related methods of use in modulating cell growth and treating cancer and cobalamin deficiency
DE602007005366D1 (de) * 2006-04-07 2010-04-29 Chimeros Inc Zusammensetzungen und verfahren zur behandlung von b-zellen-malignomen
US9044461B2 (en) 2006-04-07 2015-06-02 The Research Foundation Of State University Of New York Transcobalamin receptor polypeptides, nucleic acids, and modulators thereof, and related methods of use in modulating cell growth and treating cancer and cobalamin deficiency
WO2007120883A2 (en) * 2006-04-12 2007-10-25 Isis Pharmaceuticals, Inc. Compositions and their uses directed to hepcidin
US20100055116A1 (en) * 2006-04-13 2010-03-04 Liou Hsiou-Chi Methods and Compositions for Targeting c-Rel
HUE035732T2 (en) 2006-04-14 2018-05-28 Cell Signaling Technology Inc Gene defects and mutant ALK kinase in human solid tumors
EP2012815A4 (en) * 2006-04-14 2009-12-09 Massachusetts Inst Technology IDENTIFICATION AND MODULATION OF MOLECULAR PATHWAYS THAT INDUCE THE PLASTICITY OF THE NERVOUS SYSTEM
US7691824B2 (en) * 2006-04-28 2010-04-06 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of a gene from the JC virus
GB0608838D0 (en) 2006-05-04 2006-06-14 Novartis Ag Organic compounds
AU2007249329C1 (en) 2006-05-11 2011-03-24 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of the PCSK9 gene
WO2007133758A1 (en) * 2006-05-15 2007-11-22 Physical Pharmaceutica, Llc Composition and improved method for preparation of small particles
US8367113B2 (en) 2006-05-15 2013-02-05 Massachusetts Institute Of Technology Polymers for functional particles
US20070269892A1 (en) * 2006-05-18 2007-11-22 Nastech Pharmaceutical Company Inc. FORMULATIONS FOR INTRACELLULAR DELIVERY dsRNA
WO2007137156A2 (en) * 2006-05-19 2007-11-29 Alnylam Pharmaceuticals, Inc. Rnai modulation of aha and therapeutic uses thereof
WO2007137237A2 (en) * 2006-05-19 2007-11-29 The Scripps Research Institute Treatment of protein misfolding
WO2007137220A2 (en) * 2006-05-22 2007-11-29 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of ikk-b gene
US9273356B2 (en) 2006-05-24 2016-03-01 Medtronic, Inc. Methods and kits for linking polymorphic sequences to expanded repeat mutations
US20070275923A1 (en) * 2006-05-25 2007-11-29 Nastech Pharmaceutical Company Inc. CATIONIC PEPTIDES FOR siRNA INTRACELLULAR DELIVERY
GB0610542D0 (en) * 2006-05-26 2006-07-05 Medical Res Council Screening method
US8598333B2 (en) * 2006-05-26 2013-12-03 Alnylam Pharmaceuticals, Inc. SiRNA silencing of genes expressed in cancer
US7915399B2 (en) 2006-06-09 2011-03-29 Protiva Biotherapeutics, Inc. Modified siRNA molecules and uses thereof
EP2026843A4 (en) 2006-06-09 2011-06-22 Quark Pharmaceuticals Inc THERAPEUTIC USES OF RTP801L INHIBITORS
WO2007146953A2 (en) * 2006-06-12 2007-12-21 Exegenics, Inc., D/B/A Opko Health, Inc. Compositions and methods for sirna inhibition of angiogenesis
US9200275B2 (en) * 2006-06-14 2015-12-01 Merck Sharp & Dohme Corp. Methods and compositions for regulating cell cycle progression
WO2007150030A2 (en) 2006-06-23 2007-12-27 Massachusetts Institute Of Technology Microfluidic synthesis of organic nanoparticles
WO2008008719A2 (en) * 2006-07-10 2008-01-17 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of the myc gene
GB0613753D0 (en) * 2006-07-11 2006-08-23 Norwegian Radium Hospital Res Method
EP2447275B1 (en) 2006-07-13 2015-04-01 University of Iowa Research Foundation Methods and reagents for treatment of age-related macular degeneration
JP4756271B2 (ja) * 2006-07-18 2011-08-24 独立行政法人産業技術総合研究所 ガン細胞の老化、アポトーシス誘導剤
WO2008009477A2 (en) * 2006-07-21 2008-01-24 Silence Therapeutics Ag Means for inhibiting the expression of protein kinase 3
US20080039415A1 (en) * 2006-08-11 2008-02-14 Gregory Robert Stewart Retrograde transport of sirna and therapeutic uses to treat neurologic disorders
WO2008024844A2 (en) * 2006-08-22 2008-02-28 The Johns Hopkins University Anticancer combination therapies
DE102006039479A1 (de) 2006-08-23 2008-03-06 Febit Biotech Gmbh Programmierbare Oligonukleotidsynthese
US7872118B2 (en) * 2006-09-08 2011-01-18 Opko Ophthalmics, Llc siRNA and methods of manufacture
EP2076599A2 (en) * 2006-09-19 2009-07-08 Asuragen, Inc. Mir-200 regulated genes and pathways as targets for therapeutic intervention
WO2008036776A2 (en) * 2006-09-19 2008-03-27 Asuragen, Inc. Mir-15, mir-26, mir -31,mir -145, mir-147, mir-188, mir-215, mir-216 mir-331, mmu-mir-292-3p regulated genes and pathways as targets for therapeutic intervention
WO2008036933A2 (en) 2006-09-21 2008-03-27 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of the hamp gene
CA2663601C (en) 2006-09-22 2014-11-25 Dharmacon, Inc. Duplex oligonucleotide complexes and methods for gene silencing by rna interference
US20110150897A1 (en) * 2006-10-11 2011-06-23 Meyer Thomas F Influenza targets
WO2008063760A2 (en) * 2006-10-18 2008-05-29 The University Of Texas M.D. Anderson Cancer Center Methods for treating cancer targeting transglutaminase
JP2010507387A (ja) 2006-10-25 2010-03-11 クアーク・ファーマスーティカルス、インコーポレイテッド 新規のsiRNAおよびその使用方法
EP2061901A2 (en) 2006-10-31 2009-05-27 Noxxon Pharma AG Methods for detection of a single- or double-stranded nucleic acid molecule
EP2078079B1 (en) 2006-11-01 2011-05-04 The Medical Research and Infrastructure Fund of the Tel-Aviv Sourasky Medical Center Adipocyte-specific constructs and methods for inhibiting platelet-type 12 lipoxygenase expression
US9375440B2 (en) 2006-11-03 2016-06-28 Medtronic, Inc. Compositions and methods for making therapies delivered by viral vectors reversible for safety and allele-specificity
US8324367B2 (en) 2006-11-03 2012-12-04 Medtronic, Inc. Compositions and methods for making therapies delivered by viral vectors reversible for safety and allele-specificity
US8252526B2 (en) 2006-11-09 2012-08-28 Gradalis, Inc. ShRNA molecules and methods of use thereof
US8906874B2 (en) 2006-11-09 2014-12-09 Gradalis, Inc. Bi-functional shRNA targeting Stathmin 1 and uses thereof
US8758998B2 (en) 2006-11-09 2014-06-24 Gradalis, Inc. Construction of bifunctional short hairpin RNA
US8034921B2 (en) * 2006-11-21 2011-10-11 Alnylam Pharmaceuticals, Inc. IRNA agents targeting CCR5 expressing cells and uses thereof
US7988668B2 (en) 2006-11-21 2011-08-02 Medtronic, Inc. Microsyringe for pre-packaged delivery of pharmaceuticals
US7819842B2 (en) 2006-11-21 2010-10-26 Medtronic, Inc. Chronically implantable guide tube for repeated intermittent delivery of materials or fluids to targeted tissue sites
JP5271715B2 (ja) * 2006-11-22 2013-08-21 国立大学法人 東京大学 ジスルフィド架橋高分子ミセルを用いた環境応答性siRNAキャリア
CA2683287A1 (en) 2006-11-27 2008-12-18 Patrys Limited Novel glycosylated peptide target in neoplastic cells
JP5391073B2 (ja) 2006-11-27 2014-01-15 ディアデクサス インコーポレーテッド Ovr110抗体組成物および使用方法
WO2008067373A2 (en) * 2006-11-28 2008-06-05 Alcon Research, Ltd. RNAi-MEDIATED INHIBITION OF AQUAPORIN 1 FOR TREATMENT OF IOP-RELATED CONDITIONS
WO2008067526A2 (en) * 2006-11-30 2008-06-05 University Of Southern California Usc Stevens Compositions and methods of sphingosine kinase inhibitors in radiation therapy of various cancers
AU2007333109A1 (en) * 2006-12-08 2008-06-19 Asuragen, Inc. Functions and targets of let-7 micro RNAs
WO2008073920A2 (en) * 2006-12-08 2008-06-19 Asuragen, Inc. Mir-21 regulated genes and pathways as targets for therapeutic intervention
JP2010512747A (ja) * 2006-12-14 2010-04-30 ノバルティス アーゲー 筋疾患および心臓血管障害を処置するための組成物および方法
CA2671270A1 (en) * 2006-12-29 2008-07-17 Asuragen, Inc. Mir-16 regulated genes and pathways as targets for therapeutic intervention
US7754698B2 (en) * 2007-01-09 2010-07-13 Isis Pharmaceuticals, Inc. Modulation of FR-alpha expression
US9896511B2 (en) 2007-01-10 2018-02-20 The United States Of America, As Represented By The Secretary, Dept. Of Health And Human Services Antibodies that bind to TL1A and methods of treating inflammatory or autoimmune disease comprising administering such antibodies
US20080171906A1 (en) * 2007-01-16 2008-07-17 Everaerts Frank J L Tissue performance via hydrolysis and cross-linking
US20090074785A1 (en) * 2007-01-16 2009-03-19 Smith Jeffrey W Compositions and methods for treatment of colorectal cancer
CA2925983A1 (en) 2007-01-16 2008-07-24 The University Of Queensland Method of inducing an immune response
EP2111450A2 (en) * 2007-01-16 2009-10-28 Yissum Research Development Company of the Hebrew University of Jerusalem Nucleic acid constructs and methods for specific silencing of h19
CN102099367A (zh) * 2007-01-17 2011-06-15 蒙特利尔临床研究所 具有季碳中心的核苷和核苷酸类似物以及其使用方法
CN101641010A (zh) 2007-01-26 2010-02-03 路易斯维尔大学研究基金会公司 用作疫苗的外来体组分的修饰
WO2008093331A1 (en) * 2007-01-29 2008-08-07 Yissum Research Development Company Of The Hebrew University Of Jerusalem Antibody conjugates for circumventing multi-drug resistance
US20100183696A1 (en) * 2007-01-30 2010-07-22 Allergan, Inc Treating Ocular Diseases Using Peroxisome Proliferator-Activated Receptor Delta Antagonists
WO2008098165A2 (en) 2007-02-09 2008-08-14 Massachusetts Institute Of Technology Oscillating cell culture bioreactor
EP2121987B1 (en) 2007-02-09 2012-06-13 Northwestern University Particles for detecting intracellular targets
DE102007008596B4 (de) 2007-02-15 2010-09-02 Friedrich-Schiller-Universität Jena Biologisch wirksame Moleküle auf Grundlage von PNA und siRNA, Verfahren zu deren zellspezifischen Aktivierung sowie Applikationskit zur Verabreichung
AU2008218813B2 (en) 2007-02-20 2014-04-17 Monsanto Technology, Llc Invertebrate microRNAs
US7872119B2 (en) 2007-02-26 2011-01-18 Quark Pharmaceuticals, Inc. Inhibitors of RTP801 and their use in disease treatment
WO2008104978A2 (en) * 2007-02-28 2008-09-04 Quark Pharmaceuticals, Inc. Novel sirna structures
WO2008109357A1 (en) * 2007-03-02 2008-09-12 Mdrna, Inc. Nucleic acid compounds for inhibiting apob gene expression and uses thereof
WO2008109034A2 (en) * 2007-03-02 2008-09-12 The Trustees Of The University Of Pennsylvania Modulating pdx-1 with pcif1, methods and uses thereof
JP2010519913A (ja) * 2007-03-02 2010-06-10 エムディーアールエヌエー,インコーポレイテッド Wnt遺伝子の発現を抑制するための核酸化合物およびその使用
US9085638B2 (en) 2007-03-07 2015-07-21 The Johns Hopkins University DNA vaccine enhancement with MHC class II activators
US20080260765A1 (en) * 2007-03-15 2008-10-23 Johns Hopkins University HPV DNA Vaccines and Methods of Use Thereof
US7812002B2 (en) 2007-03-21 2010-10-12 Quark Pharmaceuticals, Inc. Oligoribonucleotide inhibitors of NRF2 and methods of use thereof for treatment of cancer
US8796442B2 (en) 2007-03-21 2014-08-05 Brookhaven Science Associates, Llc. Combined hairpin-antisense compositions and methods for modulating expression
PE20090064A1 (es) * 2007-03-26 2009-03-02 Novartis Ag Acido ribonucleico de doble cadena para inhibir la expresion del gen e6ap humano y composicion farmaceutica que lo comprende
AP2014007971A0 (en) 2007-03-29 2014-09-30 Alnylam Pharmaceuticals Inc Compositions and methods for inhibiting expressionof a gene from the ebola
WO2008124639A2 (en) * 2007-04-04 2008-10-16 Massachusetts Institute Of Technology Poly (amino acid) targeting moieties
WO2008124634A1 (en) 2007-04-04 2008-10-16 Massachusetts Institute Of Technology Polymer-encapsulated reverse micelles
CA2683063A1 (en) * 2007-04-09 2008-10-16 Chimeros, Inc. Self-assembling nanoparticle drug delivery system
CA2678055C (en) 2007-04-10 2016-02-16 Qiagen Gmbh Rna interference tags
US8808747B2 (en) * 2007-04-17 2014-08-19 Baxter International Inc. Nucleic acid microparticles for pulmonary delivery
WO2008131419A2 (en) * 2007-04-23 2008-10-30 Alnylam Pharmaceuticals, Inc. Glycoconjugates of rna interference agents
WO2008143774A2 (en) * 2007-05-01 2008-11-27 University Of Massachusetts Methods and compositions for locating snp heterozygosity for allele specific diagnosis and therapy
EP2607477B1 (en) 2007-05-03 2020-09-23 The Brigham and Women's Hospital, Inc. Multipotent stem cells and uses thereof
JP5296328B2 (ja) * 2007-05-09 2013-09-25 独立行政法人理化学研究所 1本鎖環状rnaおよびその製造方法
US20090053140A1 (en) * 2007-05-15 2009-02-26 Roderick Euan Milne Scott METHODS OF IDENTIFYING GENES INVOLVED IN MEMORY FORMATION USING SMALL INTERFERING RNA(siRNA)
KR20100029079A (ko) * 2007-05-15 2010-03-15 헬리콘 테라퓨틱스 인코퍼레이티드 Gpr12의 저해로 인지 질환을 치료하는 방법
WO2008147824A2 (en) 2007-05-22 2008-12-04 Mdrna, Inc. Hydroxymethyl substituted rna oligonucleotides and rna complexes
US20090131354A1 (en) * 2007-05-22 2009-05-21 Bader Andreas G miR-126 REGULATED GENES AND PATHWAYS AS TARGETS FOR THERAPEUTIC INTERVENTION
EP2160464B1 (en) 2007-05-30 2014-05-21 Northwestern University Nucleic acid functionalized nanoparticles for therapeutic applications
CN101820757A (zh) 2007-06-01 2010-09-01 普林斯顿大学托管委员会 通过调节宿主细胞代谢途径治疗病毒感染
KR101363928B1 (ko) * 2007-06-11 2014-02-19 고쿠리츠다이가쿠호진 미에다이가쿠 특이적 유전자 발현 방법
US20100273854A1 (en) * 2007-06-15 2010-10-28 Hagar Kalinski Compositions and methods for inhibiting nadph oxidase expression
AR066984A1 (es) * 2007-06-15 2009-09-23 Novartis Ag Inhibicion de la expresion de la subunidad alfa del canal epitelial de sodio (enac) por medio de arni (arn de interferencia)
US8785408B2 (en) 2007-06-27 2014-07-22 Quark Pharmaceuticals, Inc. Compositions and methods for reducing or protecting against delayed graft function (DGF)
EP3034095B1 (en) * 2007-06-29 2018-08-08 Stelic Institute Of Regenerative Medicine, Stelic Institute & Co. Method of fixing and expressing physiologically active substance
CN103215269B (zh) * 2007-07-05 2015-01-21 诺瓦提斯公司 用于治疗病毒感染的dsRNA
CN101821407B (zh) 2007-07-10 2013-09-18 纽瑞姆制药(1991)有限公司 神经退行性疾病中的cd44剪接变体
US8828960B2 (en) * 2007-07-17 2014-09-09 Idexx Laboratories, Inc. Amino acid vitamin ester compositions for controlled delivery of pharmaceutically active compounds
JP2009033986A (ja) * 2007-07-31 2009-02-19 Sumitomo Chemical Co Ltd RNA干渉による遺伝子発現抑制のためのターゲット遺伝子としてのcar遺伝子の使用
EP2030615A3 (en) 2007-08-13 2009-12-02 ELFORD, Howard L. Ribonucleotide reductase inhibitors for use in the treatment or prevention of neuroinflammatory or autoimmune diseases
US8501929B2 (en) * 2007-08-17 2013-08-06 Biochrom Pharma Inc. PTHrP, its isoforms and antagonist thereto in the diagnosis and treatment of disease
CA2735166C (en) 2007-08-27 2020-12-01 Boston Biomedical, Inc. Compositions of asymmetric interfering rna and uses thereof
WO2009026723A1 (en) * 2007-08-30 2009-03-05 Virexx Medical Corp. Antigenic compositions and use of same in the targeted delivery of nucleic acids
US20090081789A1 (en) * 2007-08-31 2009-03-26 Greenville Hospital System Activation of nuclear factor kappa B
WO2009033027A2 (en) 2007-09-05 2009-03-12 Medtronic, Inc. Suppression of scn9a gene expression and/or function for the treatment of pain
WO2009036368A2 (en) * 2007-09-14 2009-03-19 Nitto Denko Corporation Drug carriers
JP5049713B2 (ja) * 2007-09-14 2012-10-17 株式会社コナミデジタルエンタテインメント ゲームシステム並びにこれを構成するゲーム装置及び課題報知装置
US8361714B2 (en) 2007-09-14 2013-01-29 Asuragen, Inc. Micrornas differentially expressed in cervical cancer and uses thereof
EP2195428B1 (en) 2007-09-19 2013-12-11 Applied Biosystems, LLC SiRNA SEQUENCE-INDEPENDENT MODIFICATION FORMATS FOR REDUCING OFF-TARGET PHENOTYPIC EFFECTS IN RNAi, AND STABILIZED FORMS THEREOF
EP2197454A4 (en) * 2007-09-25 2012-07-04 Idexx Lab Inc PHARMACEUTICAL COMPOSITIONS FOR THE ADMINISTRATION OF OLIGONUCLEOTIDES
ES2376175T3 (es) 2007-09-28 2012-03-09 Bind Biosciences, Inc. Direccionamiento a células de c�?ncer usando nanopart�?culas.
US20120082659A1 (en) * 2007-10-02 2012-04-05 Hartmut Land Methods And Compositions Related To Synergistic Responses To Oncogenic Mutations
JP5646997B2 (ja) 2007-10-03 2014-12-24 クォーク ファーマシューティカルズ インコーポレーティッドQuark Pharmaceuticals,Inc. 新規siRNA構造
JP2010539990A (ja) * 2007-10-04 2010-12-24 ボード オブ リージェンツ ザ ユニバーシティー オブ テキサス システム アンチセンス転写物を標的とするagRNAおよびギャップマーを用いた遺伝子発現の調節方法
AU2008314647B2 (en) 2007-10-12 2013-03-21 Massachusetts Institute Of Technology Vaccine nanotechnology
US9096855B2 (en) 2007-10-18 2015-08-04 Cell Signaling Technology, Inc. Translocation and mutant ROS kinase in human non-small cell lung carcinoma
US8097712B2 (en) * 2007-11-07 2012-01-17 Beelogics Inc. Compositions for conferring tolerance to viral disease in social insects, and the use thereof
US20100098664A1 (en) * 2007-11-28 2010-04-22 Mathieu Jean-Francois Desclaux Lentiviral vectors allowing RNAi mediated inhibition of GFAP and vimentin expression
US8324369B2 (en) * 2007-11-30 2012-12-04 Baylor College Of Medicine Dendritic cell vaccine compositions and uses of same
US8071562B2 (en) 2007-12-01 2011-12-06 Mirna Therapeutics, Inc. MiR-124 regulated genes and pathways as targets for therapeutic intervention
EP2268664B1 (en) 2007-12-03 2017-05-24 The Government of the United States of America as represented by the Secretary of the Department of Health and Human Services Doc1 compositions and methods for treating cancer
EP4074344A1 (en) 2007-12-04 2022-10-19 Arbutus Biopharma Corporation Targeting lipids
AU2008340354B2 (en) * 2007-12-04 2014-04-17 Alnylam Pharmaceuticals, Inc. Folate-iRNA conjugates
WO2009076400A2 (en) 2007-12-10 2009-06-18 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of factor vii gene
US8614311B2 (en) 2007-12-12 2013-12-24 Quark Pharmaceuticals, Inc. RTP801L siRNA compounds and methods of use thereof
US20110105584A1 (en) * 2007-12-12 2011-05-05 Elena Feinstein Rtp80il sirna compounds and methods of use thereof
EP2229459B1 (en) 2007-12-13 2014-08-27 Alnylam Pharmaceuticals, Inc. Methods and compositions for prevention or treatment of RSV infection
WO2009079399A2 (en) * 2007-12-14 2009-06-25 Alnylam Pharmaceuticals, Inc. Method of treating neurodegenerative disease
US7845686B2 (en) * 2007-12-17 2010-12-07 S & B Technical Products, Inc. Restrained pipe joining system for plastic pipe
KR100949791B1 (ko) * 2007-12-18 2010-03-30 이동기 오프-타겟 효과를 최소화하고 RNAi 기구를 포화시키지않는 신규한 siRNA 구조 및 그 용도
WO2009086156A2 (en) * 2007-12-21 2009-07-09 Asuragen, Inc. Mir-10 regulated genes and pathways as targets for therapeutic intervention
EP2242854A4 (en) * 2008-01-15 2012-08-15 Quark Pharmaceuticals Inc COMPOUNDS AND USES THEREOF
AU2009241591A1 (en) * 2008-01-31 2009-11-05 Alnylam Pharmaceuticals, Inc. Optimized methods for delivery of DSRNA targeting the PCSK9 gene
US20090263803A1 (en) * 2008-02-08 2009-10-22 Sylvie Beaudenon Mirnas differentially expressed in lymph nodes from cancer patients
US8188060B2 (en) 2008-02-11 2012-05-29 Dharmacon, Inc. Duplex oligonucleotides with enhanced functionality in gene regulation
US7977321B2 (en) * 2008-02-12 2011-07-12 University Of Tennessee Research Foundation Small interfering RNAs targeting feline herpes virus
EP2250266A2 (en) 2008-02-12 2010-11-17 Alnylam Pharmaceuticals Inc. Compositions and methods for inhibiting expression of cd45 gene
DE102009043743B4 (de) 2009-03-13 2016-10-13 Friedrich-Schiller-Universität Jena Zellspezifisch wirksame Moleküle auf Grundlage von siRNA sowie Applikationskits zu deren Herstellung und Verwendung
EP2247748A2 (en) * 2008-02-13 2010-11-10 Elan Pharma International Limited Alpha-synuclein kinase
WO2009103067A2 (en) * 2008-02-14 2009-08-20 The Children's Hospital Of Philadelphia Compositions and methods to treat asthma
WO2009111658A2 (en) * 2008-03-05 2009-09-11 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of eg5 and vegf genes
WO2009111643A2 (en) * 2008-03-06 2009-09-11 Asuragen, Inc. Microrna markers for recurrence of colorectal cancer
US8202848B2 (en) * 2008-03-17 2012-06-19 Board Of Regents, The University Of Texas System Identification of micro-RNAS involved in neuromuscular synapse maintenance and regeneration
WO2009116037A2 (en) * 2008-03-20 2009-09-24 Quark Pharmaceuticals, Inc. NOVEL siRNA COMPOUNDS FOR INHIBITING RTP801
US20090253780A1 (en) * 2008-03-26 2009-10-08 Fumitaka Takeshita COMPOSITIONS AND METHODS RELATED TO miR-16 AND THERAPY OF PROSTATE CANCER
EP2105145A1 (en) * 2008-03-27 2009-09-30 ETH Zürich Method for muscle-specific delivery lipid-conjugated oligonucleotides
WO2009123185A1 (ja) * 2008-03-31 2009-10-08 独立行政法人産業技術総合研究所 Rna干渉効果が高い2本鎖脂質修飾rna
TWI348916B (en) * 2008-04-03 2011-09-21 Univ Nat Taiwan A novel treatment tool for cancer: rna interference of bcas2
US20090258928A1 (en) * 2008-04-08 2009-10-15 Asuragen, Inc. Methods and compositions for diagnosing and modulating human papillomavirus (hpv)
AU2009234266B2 (en) 2008-04-11 2015-08-06 Tekmira Pharmaceuticals Corporation Site-specific delivery of nucleic acids by combining targeting ligands with endosomolytic components
US8309614B2 (en) * 2008-04-11 2012-11-13 Cedars-Sinai Medical Center Poly(beta malic acid) with pendant leu-leu-leu tripeptide for effective cytoplasmic drug delivery
WO2009127060A1 (en) 2008-04-15 2009-10-22 Protiva Biotherapeutics, Inc. Novel lipid formulations for nucleic acid delivery
US8278287B2 (en) * 2008-04-15 2012-10-02 Quark Pharmaceuticals Inc. siRNA compounds for inhibiting NRF2
US20090285861A1 (en) * 2008-04-17 2009-11-19 Tzyy-Choou Wu Tumor cell-based cancer immunotherapeutic compositions and methods
US7875711B2 (en) * 2008-04-17 2011-01-25 Alnylam Pharamaceuticals, Inc. Compositions and methods for inhibiting expression of XBP-1 gene
USRE48948E1 (en) 2008-04-18 2022-03-01 Warsaw Orthopedic, Inc. Clonidine compounds in a biodegradable polymer
US9005599B2 (en) 2008-04-21 2015-04-14 Tissue Regeneration Therapeutics Inc. Genetically modified human umbilical cord perivascular cells for prophylaxis against or treatment of biological or chemical agents
US8324366B2 (en) 2008-04-29 2012-12-04 Alnylam Pharmaceuticals, Inc. Compositions and methods for delivering RNAI using lipoproteins
WO2009134443A2 (en) * 2008-05-02 2009-11-05 The Brigham And Women's Hospital, Inc. Rna-induced translational silencing and cellular apoptosis
WO2009137807A2 (en) 2008-05-08 2009-11-12 Asuragen, Inc. Compositions and methods related to mirna modulation of neovascularization or angiogenesis
US20090291073A1 (en) * 2008-05-20 2009-11-26 Ward Keith W Compositions Comprising PKC-theta and Methods for Treating or Controlling Ophthalmic Disorders Using Same
US20100009451A1 (en) * 2008-05-30 2010-01-14 Sigma Aldrich Company Compositions and methods for specifically silencing a target nucleic acid
EP2297322A1 (en) 2008-06-04 2011-03-23 The Board of Regents of The University of Texas System Modulation of gene expression through endogenous small rna targeting of gene promoters
US20090305611A1 (en) * 2008-06-06 2009-12-10 Flow International Corporation Device and method for improving accuracy of a high-pressure fluid jet apparatus
JP5524189B2 (ja) * 2008-06-06 2014-06-18 クォーク ファーマシューティカルズ インコーポレーティッド 耳障害治療のための組成物および方法
EP2235177B1 (en) * 2008-06-13 2012-07-18 RiboxX GmbH Method for enzymatic synthesis of chemically modified rna
WO2010005741A1 (en) * 2008-06-16 2010-01-14 Georgia Tech Research Corporation Nanogels for cellular delivery of therapeutics
TWI455944B (zh) 2008-07-01 2014-10-11 Daiichi Sankyo Co Ltd 雙股多核苷酸
WO2010006342A2 (en) 2008-07-11 2010-01-14 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of gsk-3 genes
US8309791B2 (en) 2008-07-16 2012-11-13 Recombinectics, Inc. Method for producing a transgenic pig using a hyper-methylated transposon
US8039658B2 (en) * 2008-07-25 2011-10-18 Air Products And Chemicals, Inc. Removal of trace arsenic impurities from triethylphosphate (TEPO)
WO2010014857A2 (en) * 2008-07-30 2010-02-04 University Of Massachusetts Chromosome therapy
EP2326351B1 (en) 2008-08-19 2017-12-27 Nektar Therapeutics Conjugates of small-interfering nucleic acids
AU2009275387B2 (en) 2008-08-25 2010-07-08 Excaliard Pharmaceuticals, Inc. Antisense oligonucleotides directed against connective tissue growth factor and uses thereof
WO2011028218A1 (en) 2009-09-02 2011-03-10 Alnylam Pharmaceuticals, Inc. Process for triphosphate oligonucleotide synthesis
EP3208339B1 (en) 2008-09-15 2019-05-01 The Children's Medical Center Corporation Modulation of bcl11a for treatment of hemoglobinopathies
JP2012513953A (ja) 2008-09-23 2012-06-21 アルニラム ファーマスーティカルズ インコーポレイテッド 付加環化を用いたモノマーおよびオリゴヌクレオチドの化学修飾
JP5529142B2 (ja) 2008-09-25 2014-06-25 アルナイラム ファーマシューティカルズ, インコーポレイテッド 血清アミロイドa遺伝子の発現を阻害するための脂質製剤組成物および方法
US8592570B2 (en) 2008-10-06 2013-11-26 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of an RNA from West Nile virus
US9149492B2 (en) 2008-10-08 2015-10-06 Trustees Of Dartmouth College Method for selectively inhibiting ACAT1 in the treatment of alzheimer's disease
US8802646B2 (en) * 2008-10-08 2014-08-12 Trustees Of Dartmouth College Method for selectively inhibiting the activity of ACAT1 in the treatment of alzheimer's disease
US9388413B2 (en) 2008-10-08 2016-07-12 Trustees Of Dartmouth College Method for selectively inhibiting ACAT1 in the treatment of neurodegenerative diseases
US9388414B2 (en) 2008-10-08 2016-07-12 Trustees Of Dartmouth College Method for selectively inhibiting ACAT1 in the treatment of neurodegenerative diseases
WO2010042292A1 (en) * 2008-10-08 2010-04-15 Trustees Of Dartmouth College Method for selectively inhibiting the activity of acat1 in the treatment of alzheimer's disease
EP2350043B9 (en) 2008-10-09 2014-08-20 TEKMIRA Pharmaceuticals Corporation Improved amino lipids and methods for the delivery of nucleic acids
US8591905B2 (en) 2008-10-12 2013-11-26 The Brigham And Women's Hospital, Inc. Nicotine immunonanotherapeutics
US8343497B2 (en) 2008-10-12 2013-01-01 The Brigham And Women's Hospital, Inc. Targeting of antigen presenting cells with immunonanotherapeutics
US8343498B2 (en) 2008-10-12 2013-01-01 Massachusetts Institute Of Technology Adjuvant incorporation in immunonanotherapeutics
US8277812B2 (en) 2008-10-12 2012-10-02 Massachusetts Institute Of Technology Immunonanotherapeutics that provide IgG humoral response without T-cell antigen
US9458472B2 (en) * 2008-10-15 2016-10-04 Massachusetts Institute Of Technology Detection and destruction of cancer cells using programmed genetic vectors
EP2352744B1 (en) * 2008-10-15 2016-09-21 Somagenics, Inc. Short hairpin rnas for inhibition of gene expression
US8168775B2 (en) 2008-10-20 2012-05-01 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of transthyretin
EP2350277A1 (en) * 2008-10-23 2011-08-03 Alnylam Pharmaceuticals, Inc. Methods and compositions for prevention or treatment of rsv infection using modified duplex rna molecules
WO2010046889A1 (en) * 2008-10-23 2010-04-29 Quark Pharmaceuticals, Inc. Methods for delivery of sirna to bone marrow cells and uses thereof
KR20230147211A (ko) 2008-11-10 2023-10-20 알닐람 파마슈티칼스 인코포레이티드 치료제 운반용 신규 지질 및 조성물
WO2010056737A2 (en) * 2008-11-11 2010-05-20 Mirna Therapeutics, Inc. Methods and compositions involving mirnas in cancer stem cells
EP2365804B1 (en) * 2008-11-13 2015-05-06 Modgene, Llc Reduction of amyloid-beta load in non-brain tissue
JP2012509331A (ja) 2008-11-21 2012-04-19 アイシス ファーマシューティカルズ, インコーポレーテッド がんの治療のための併用療法
KR101692880B1 (ko) 2008-11-24 2017-01-04 노오쓰웨스턴 유니버시티 다가 rna-나노입자 구조체
EP2191834A1 (en) * 2008-11-26 2010-06-02 Centre National De La Recherche Scientifique (Cnrs) Compositions and methods for treating retrovirus infections
CA2744093A1 (en) 2008-12-03 2010-06-10 Marina Biotech, Inc. Una oligomer structures for therapeutic agents
US20100291188A1 (en) * 2008-12-04 2010-11-18 Musc Foundation For Research Development Periostin Inhibitory Compositions for Myocardial Regeneration, Methods of Delivery, and Methods of Using Same
MX2011005851A (es) 2008-12-04 2011-07-29 Opko Opthalmics Llc Composiciones y metodos para la inhibicion selectiva de isoformas vegf proangiogenicas.
CA2740806C (en) 2008-12-09 2021-07-20 Bryan Irving Anti-pd-l1 antibodies and their use to enhance t-cell function
JP5855462B2 (ja) 2008-12-10 2016-02-09 アルナイラム ファーマシューティカルズ, インコーポレイテッドAlnylam Pharmaceuticals, Inc. GNAQを標的としたdsRNA組成物および発現を阻害するための方法
WO2010066112A1 (en) * 2008-12-11 2010-06-17 The University Of Hong Kong siRNA COMPOSITIONS AND METHODS FOR POTENTLY INHIBITING VIRAL INFECTION
JPWO2010067882A1 (ja) * 2008-12-12 2012-05-24 株式会社クレハ 癌及び喘息治療のための医薬組成物
EP2370175A2 (en) * 2008-12-16 2011-10-05 Bristol-Myers Squibb Company Methods of inhibiting quiescent tumor proliferation
US20110288155A1 (en) 2008-12-18 2011-11-24 Elena Feinstein Sirna compounds and methods of use thereof
US20110268772A1 (en) 2008-12-26 2011-11-03 Samyang Corporation Pharmaceutical composition containing an anionic drug and a production method thereof
US20100233270A1 (en) 2009-01-08 2010-09-16 Northwestern University Delivery of Oligonucleotide-Functionalized Nanoparticles
WO2010083532A1 (en) * 2009-01-19 2010-07-22 The Research Foundaton Of State University Of New York Fatty acid binding proteins as drug targets for modulation of endocannabinoids
EP2389196A1 (en) * 2009-01-20 2011-11-30 Vib Vzw Phd2 inhibition for blood vessel normalization, and uses thereof
WO2010083615A1 (en) 2009-01-26 2010-07-29 Protiva Biotherapeutics, Inc. Compositions and methods for silencing apolipoprotein c-iii expression
AU2010211133A1 (en) * 2009-02-03 2011-07-21 F. Hoffmann-La Roche Ag Compositions and methods for inhibiting expression of PTP1B genes
EP2881402B1 (en) 2009-02-12 2017-05-10 Cell Signaling Technology, Inc. Mutant ROS expression in human liver cancer
US8975389B2 (en) 2009-03-02 2015-03-10 Alnylam Pharmaceuticals, Inc. Nucleic acid chemical modifications
MX2011009724A (es) * 2009-03-19 2011-10-14 Merck Sharp & Dohme Inhibicion mediada por interferencia de acido ribonucleico de la expresion del gen del factor de transcripcion basico de cierre de leucina 1 de homologia de btb y cnc 1, usando el listado de secuencias de acido nucleico corto de interferencia.
US8444983B2 (en) 2009-03-23 2013-05-21 Quark Pharmaceuticals, Inc. Composition of anti-ENDO180 antibodies and methods of use for the treatment of cancer and fibrotic diseases
US20100239632A1 (en) 2009-03-23 2010-09-23 Warsaw Orthopedic, Inc. Drug depots for treatment of pain and inflammation in sinus and nasal cavities or cardiac tissue
WO2010120874A2 (en) 2009-04-14 2010-10-21 Chimeros, Inc. Chimeric therapeutics, compositions, and methods for using same
EP2421972A2 (en) * 2009-04-24 2012-02-29 The Board of Regents of The University of Texas System Modulation of gene expression using oligomers that target gene regions downstream of 3' untranslated regions
US8367350B2 (en) 2009-04-29 2013-02-05 Morehouse School Of Medicine Compositions and methods for diagnosis, prognosis and management of malaria
US8933049B2 (en) * 2009-05-05 2015-01-13 Medical Diagnostic Laboratories, Llc Repressor on IFN-λ promoter and siRNA against ZEB1 and BLIMP-1 to increase IFN-λ gene activity
AU2010244122B2 (en) 2009-05-05 2015-05-14 Beeologics Inc. Prevention and treatment of Nosema disease in bees
US9255266B2 (en) * 2009-05-06 2016-02-09 Rutgers, The State University Of New Jersey RNA targeting in alpha-synucleinopathies
EP2429657A2 (en) * 2009-05-15 2012-03-21 F. Hoffmann-La Roche AG Compositions and methods for inhibiting expression of glucocorticoid receptor (gcr) genes
WO2011005363A2 (en) * 2009-05-18 2011-01-13 Ensysce Biosciences, Inc. Carbon nanotubes complexed with multiple bioactive agents and methods related thereto
WO2011019423A2 (en) 2009-05-20 2011-02-17 Schering Corporation Modulation of pilr receptors to treat microbial infections
WO2010135669A1 (en) * 2009-05-22 2010-11-25 Sabiosciences Corporation Arrays and methods for reverse genetic functional analysis
EP2438168B1 (en) 2009-06-01 2020-02-12 Halo-Bio Rnai Therapeutics, Inc. Polynucleotides for multivalent rna interference, compositions and methods of use thereof
US20120083519A1 (en) * 2009-06-03 2012-04-05 Djillali Sahali Methods For Diagnosing And Treating A Renal Disease In An Individual
EP2437592A4 (en) 2009-06-05 2013-02-27 Univ Florida ISOLATION AND TARGETED SUPPRESSION OF GENES OF LIGNIN BIOSYNTHESIS IN SUGAR CANE
CA2763537A1 (en) 2009-06-08 2010-12-16 Quark Pharmaceuticals, Inc. Methods for treating chronic kidney disease
NZ622843A (en) 2009-06-10 2015-10-30 Tekmira Pharmaceuticals Corp Improved lipid formulation
CN107129998A (zh) 2009-06-10 2017-09-05 淡马锡生命科学研究院有限公司 用于棉花中基因功能性分析的病毒诱导的基因沉默(vigs)
US8273869B2 (en) * 2009-06-15 2012-09-25 Alnylam Pharmaceuticals, Inc. Lipid formulated dsRNA targeting the PCSK9 gene
US9051567B2 (en) 2009-06-15 2015-06-09 Tekmira Pharmaceuticals Corporation Methods for increasing efficacy of lipid formulated siRNA
US20100323018A1 (en) * 2009-06-17 2010-12-23 Massachusetts Institute Of Technology Branched DNA/RNA monomers and uses thereof
US20100324124A1 (en) * 2009-06-17 2010-12-23 Massachusetts Institute Of Technology Compositions and methods relating to DNA-based particles
GB0910723D0 (en) * 2009-06-22 2009-08-05 Sylentis Sau Novel drugs for inhibition of gene expression
US9018187B2 (en) 2009-07-01 2015-04-28 Protiva Biotherapeutics, Inc. Cationic lipids and methods for the delivery of therapeutic agents
CA2767127A1 (en) 2009-07-01 2011-01-06 Protiva Biotherapeutics, Inc. Novel lipid formulations for delivery of therapeutic agents to solid tumors
US8569256B2 (en) 2009-07-01 2013-10-29 Protiva Biotherapeutics, Inc. Cationic lipids and methods for the delivery of therapeutic agents
WO2011008730A2 (en) 2009-07-13 2011-01-20 Somagenics Inc. Chemical modification of small hairpin rnas for inhibition of gene expression
US8716464B2 (en) * 2009-07-20 2014-05-06 Thomas W. Geisbert Compositions and methods for silencing Ebola virus gene expression
EP2769737B1 (en) 2009-07-20 2017-04-05 Bristol-Myers Squibb Company Combination of an anti-CTLA4 antibody with etoposide for the synergistic treatment of proliferative diseases
CA2769822C (en) 2009-08-13 2019-02-19 The Johns Hopkins University Methods of modulating immune function
AP2015008874A0 (en) 2009-08-14 2015-11-30 Alnylam Pharmaceuticals Inc Lipid formulated compositions and methods for inhibiting expression of a gene from the ebola virus
AP3412A (en) 2009-08-24 2015-09-30 Phigenix Inc Targeting pax2 for the treatment of breast cancer
WO2011031600A1 (en) 2009-09-10 2011-03-17 Schering Corporation Use of il-33 antagonists to treat fibrotic disease
WO2011032100A1 (en) 2009-09-11 2011-03-17 Government Of The U.S.A., As Represented By The Secretary, Department Of Health And Human Services Inhibitors of kshv vil6 and human il6
EP2295543A1 (en) 2009-09-11 2011-03-16 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Method for the preparation of an influenza virus
CN107519133A (zh) 2009-09-15 2017-12-29 阿尔尼拉姆医药品有限公司 脂质配制的组合物及抑制Eg5和VEGF基因的表达的方法
WO2011035065A1 (en) 2009-09-17 2011-03-24 Nektar Therapeutics Monoconjugated chitosans as delivery agents for small interfering nucleic acids
US9187746B2 (en) 2009-09-22 2015-11-17 Alnylam Pharmaceuticals, Inc. Dual targeting siRNA agents
US9222086B2 (en) * 2009-09-23 2015-12-29 Protiva Biotherapeutics, Inc. Compositions and methods for silencing genes expressed in cancer
US20150025122A1 (en) 2009-10-12 2015-01-22 Larry J. Smith Methods and Compositions for Modulating Gene Expression Using Oligonucleotide Based Drugs Administered in vivo or in vitro
US8962584B2 (en) 2009-10-14 2015-02-24 Yissum Research Development Company Of The Hebrew University Of Jerusalem, Ltd. Compositions for controlling Varroa mites in bees
AU2010313154B2 (en) 2009-10-30 2016-05-12 Northwestern University Templated nanoconjugates
WO2011056883A1 (en) 2009-11-03 2011-05-12 Alnylam Pharmaceuticals, Inc. Lipid formulated compositions and methods for inhibiting expression of transthyretin (ttr)
US9799416B2 (en) * 2009-11-06 2017-10-24 Terrapower, Llc Methods and systems for migrating fuel assemblies in a nuclear fission reactor
US8901097B2 (en) 2009-11-08 2014-12-02 Quark Pharmaceuticals, Inc. Methods for delivery of siRNA to the spinal cord and therapies arising therefrom
WO2011062997A2 (en) 2009-11-17 2011-05-26 Musc Foundation For Research Development Human monoclonal antibodies to human nucleolin
EP2504435B1 (en) 2009-11-26 2019-11-13 Quark Pharmaceuticals, Inc. Sirna compounds comprising terminal substitutions
WO2011071860A2 (en) 2009-12-07 2011-06-16 Alnylam Pharmaceuticals, Inc. Compositions for nucleic acid delivery
PL2509991T3 (pl) * 2009-12-09 2016-04-29 Nitto Denko Corp Modulacja ekspresji hsp47
US8778904B2 (en) 2009-12-09 2014-07-15 Quark Pharmaceuticals, Inc. Methods and compositions for treating diseases, disorders or injury of the CNS
WO2011072240A1 (en) 2009-12-10 2011-06-16 Cedars-Sinai Medical Center Drug delivery of temozolomide for systemic based treatment of cancer
EP2336171A1 (en) 2009-12-11 2011-06-22 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Novel targets for the treatment of proliferative diseases
EP2513308B1 (en) 2009-12-17 2017-01-18 Merck Sharp & Dohme Corp. Modulation of pilr to treat immune disorders
WO2011073326A2 (en) 2009-12-18 2011-06-23 Novartis Ag Organic compositions to treat hsf1-related diseases
US20130017223A1 (en) 2009-12-18 2013-01-17 The University Of British Columbia Methods and compositions for delivery of nucleic acids
EP2516645A1 (en) 2009-12-23 2012-10-31 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Influenza targets
KR101589951B1 (ko) 2009-12-23 2016-01-29 그래댈리스, 인코포레이티드 푸린―녹다운 및 gm―csf―증강된 (fang) 암 백신
US20130023578A1 (en) 2009-12-31 2013-01-24 Samyang Biopharmaceuticals Corporation siRNA for inhibition of c-Met expression and anticancer composition containing the same
TW201124159A (en) * 2010-01-07 2011-07-16 Univ Nat Cheng Kung Small interference RNA molecule and applications thereof
WO2011084193A1 (en) 2010-01-07 2011-07-14 Quark Pharmaceuticals, Inc. Oligonucleotide compounds comprising non-nucleotide overhangs
DK2524039T3 (en) * 2010-01-11 2018-03-12 Curna Inc TREATMENT OF GENDER HORMON-BINDING GLOBULIN (SHBG) RELATED DISEASES BY INHIBITION OF NATURAL ANTISENCE TRANSCRIPTS TO SHBG
WO2011088058A1 (en) * 2010-01-12 2011-07-21 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expressions of factor vii and pten genes
DE102010004957A1 (de) 2010-01-14 2011-07-21 Universitätsklinikum Jena, 07743 Biologisch wirksame Moleküle zur Beeinflussung von Virus-, Bakterien-, Parasiten-infizierten Zellen und/oder Tumorzellen und Verfahren zu deren Anwendung
WO2011094580A2 (en) 2010-01-28 2011-08-04 Alnylam Pharmaceuticals, Inc. Chelated copper for use in the preparation of conjugated oligonucleotides
WO2011100131A2 (en) 2010-01-28 2011-08-18 Alnylam Pharmacuticals, Inc. Monomers and oligonucleotides comprising cycloaddition adduct(s)
US8722641B2 (en) 2010-01-29 2014-05-13 St. Jude Children's Research Hospital Oligonucleotides which inhibit p53 induction in response to cellular stress
EP2534489A1 (en) 2010-02-10 2012-12-19 Novartis AG Methods and compounds for muscle growth
WO2011116351A2 (en) 2010-03-19 2011-09-22 University Of South Alabama Methods and compositions for the treatment of cancer
US8455455B1 (en) 2010-03-31 2013-06-04 Protiva Biotherapeutics, Inc. Compositions and methods for silencing genes involved in hemorrhagic fever
US9102938B2 (en) 2010-04-01 2015-08-11 Alnylam Pharmaceuticals, Inc. 2′ and 5′ modified monomers and oligonucleotides
WO2011133871A2 (en) 2010-04-22 2011-10-27 Alnylam Pharmaceuticals, Inc. 5'-end derivatives
WO2011133868A2 (en) 2010-04-22 2011-10-27 Alnylam Pharmaceuticals, Inc. Conformationally restricted dinucleotide monomers and oligonucleotides
WO2011133658A1 (en) 2010-04-22 2011-10-27 Boston Medical Center Corporation Compositions and methods for targeting and delivering therapeutics into cells
US10913767B2 (en) 2010-04-22 2021-02-09 Alnylam Pharmaceuticals, Inc. Oligonucleotides comprising acyclic and abasic nucleosides and analogs
JP5896175B2 (ja) 2010-04-29 2016-03-30 アイオーニス ファーマシューティカルズ, インコーポレーテッドIonis Pharmaceuticals,Inc. トランスサイレチン発現の調節
WO2011137363A1 (en) 2010-04-30 2011-11-03 Allergan, Inc. Novel treatment for age related macular degeneration and ocular ischemic disease associated with complement activation by targeting 5-lipoxygenase
KR20190000385A (ko) 2010-05-04 2019-01-02 더 브리검 앤드 우먼즈 하스피털, 인크. 섬유증의 검출 및 치료
US8563243B2 (en) * 2010-05-12 2013-10-22 University Of South Carolina Methods for affecting homology-directed DNA double stranded break repair
NO2575876T3 (zh) 2010-05-26 2018-05-05
EP2390327A1 (en) 2010-05-27 2011-11-30 Sylentis S.A. siRNA and their use in methods and compositions for the treatment and/or prevention of eye conditions
DE102010022937A1 (de) 2010-06-04 2011-12-08 Universitätsklinikum Jena Zellspezifisch aktivierbare biologisch wirksame Moleküle auf Grundlage von siRNA, Verfahren zu deren Aktivierung sowie Applikationskit zur Verabreichung
WO2011163121A1 (en) 2010-06-21 2011-12-29 Alnylam Pharmaceuticals, Inc. Multifunctional copolymers for nucleic acid delivery
NZ604094A (en) 2010-06-24 2014-11-28 Quark Pharmaceuticals Inc Double stranded rna compounds to rhoa and use thereof
US9006417B2 (en) 2010-06-30 2015-04-14 Protiva Biotherapeutics, Inc. Non-liposomal systems for nucleic acid delivery
WO2012016184A2 (en) 2010-07-30 2012-02-02 Alnylam Pharmaceuticals, Inc. Methods and compositions for delivery of active agents
WO2012016188A2 (en) 2010-07-30 2012-02-02 Alnylam Pharmaceuticals, Inc. Methods and compositions for delivery of active agents
US20120101108A1 (en) 2010-08-06 2012-04-26 Cell Signaling Technology, Inc. Anaplastic Lymphoma Kinase In Kidney Cancer
EP2609198B8 (en) 2010-08-24 2018-03-28 Sirna Therapeutics, Inc. SINGLE-STRANDED RNAi AGENTS CONTAINING AN INTERNAL, NON-NUCLEIC ACID SPACER
EP2622076A1 (en) 2010-09-30 2013-08-07 University of Zürich Treatment of b-cell lymphoma with microrna
SG189280A1 (en) * 2010-10-07 2013-05-31 Agency Science Tech & Res Parp-1 inhibitors
WO2012051491A1 (en) 2010-10-14 2012-04-19 The United States Of America, As Represented By The Secretary National Institutes Of Health Compositions and methods for controlling neurotropic viral pathogenesis by micro-rna targeting
EP2631291B1 (en) 2010-10-22 2019-05-15 OliX Pharmaceuticals, Inc. Nucleic acid molecules inducing rna interference, and uses thereof
US9260471B2 (en) 2010-10-29 2016-02-16 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using short interfering nucleic acids (siNA)
WO2012071436A1 (en) 2010-11-24 2012-05-31 Genentech, Inc. Method of treating autoimmune inflammatory disorders using il-23r loss-of-function mutants
US20130324591A1 (en) 2010-12-06 2013-12-05 Quark Pharmaceuticals, Inc. Double stranded oligonucleotide compounds comprising positional modifications
US10202615B2 (en) 2010-12-10 2019-02-12 Vanderbilt University Mammalian genes involved in toxicity and infection
US9617542B2 (en) * 2010-12-14 2017-04-11 The United States of America, as representd by The Secretary of Agriculture Lepidopteran moth control using double-stranded RNA constructs
US9623041B2 (en) 2010-12-30 2017-04-18 Cedars-Sinai Medical Center Polymalic acid-based nanobiopolymer compositions
KR102119187B1 (ko) 2011-01-10 2020-06-08 더 리젠츠 오브 더 유니버시티 오브 미시간 줄기 세포 인자 억제제
US20150018408A1 (en) 2013-07-10 2015-01-15 The Regents Of The University Of Michigan Therapeutic antibodies and uses thereof
EP3202760B1 (en) 2011-01-11 2019-08-21 Alnylam Pharmaceuticals, Inc. Pegylated lipids and their use for drug delivery
DE102011009470A1 (de) 2011-01-21 2012-08-09 Friedrich-Schiller-Universität Jena Biologisch wirksame Nukleotid-Moleküle zur gezielten Abtötung von Zellen, Verwendung derselben sowie Applikationskit
TWI593416B (zh) 2011-02-02 2017-08-01 艾克厘德製藥公司 利用針對結締組織生長因子(ctgf)目標之反義化合物治療瘢痕或肥厚性疤痕之方法
US9222085B2 (en) 2011-02-03 2015-12-29 Mirna Therapeutics, Inc. Synthetic mimics of MIR-124
SG193280A1 (en) * 2011-03-03 2013-10-30 Quark Pharmaceuticals Inc Oligonucleotide modulators of the toll-like receptor pathway
US9796979B2 (en) 2011-03-03 2017-10-24 Quark Pharmaceuticals Inc. Oligonucleotide modulators of the toll-like receptor pathway
WO2012125554A2 (en) * 2011-03-11 2012-09-20 Board Of Regents Of The University Of Nebraska Compositions and methods for the treatment of cancer
JP2014510526A (ja) 2011-03-15 2014-05-01 ユニヴァーシティー オブ ユタ リサーチ ファウンデーション 血管関連黄斑症およびその症状の診断および治療方法
WO2012135696A2 (en) * 2011-04-01 2012-10-04 University Of South Alabama Methods and compositions for the diagnosis, classification, and treatment of cancer
US10266565B2 (en) 2011-04-12 2019-04-23 The United States Of America, As Represented By The Secretary, Department Of Health & Human Services Peptide mimetic ligands of polo-like kinase 1 polo box domain and methods of use
ES2634450T3 (es) 2011-04-13 2017-09-27 Ionis Pharmaceuticals, Inc. Modulación antisentido de la expresión de PTP1B
CA2833269C (en) * 2011-04-15 2020-04-14 Molecular Transfer, Inc. Agents for improved delivery of nucleic acids to eukaryotic cells
US8716257B2 (en) * 2011-04-15 2014-05-06 Sutter West Bay Hospitals CMV gene products promote cancer stem cell growth
KR20140104344A (ko) 2011-05-20 2014-08-28 더 유나이티드 스테이츠 오브 어메리카, 애즈 리프리젠티드 바이 더 시크리터리, 디파트먼트 오브 헬쓰 앤드 휴먼 서비시스 T 세포 매개성 질병의 병증을 개선하기 위한 tl1a-dr3의 상호작용의 차단 및 그것의 항체
US9452219B2 (en) 2011-06-02 2016-09-27 University Of Louisville Research Foundation, Inc. Anti-nucleolin agent-conjugated nanoparticles
MX344807B (es) 2011-06-21 2017-01-09 Alnylam Pharmaceuticals Inc Composiciones y metodos para inhibicion de genes para alipoproteina c-iii (apoc3).
US9228188B2 (en) 2011-06-21 2016-01-05 Alnylam Pharmaceuticals, Inc. Compositions and method for inhibiting hepcidin antimicrobial peptide (HAMP) or HAMP-related gene expression
CN112961855A (zh) 2011-06-21 2021-06-15 阿尔尼拉姆医药品有限公司 血管生成素样3(ANGPTL3)iRNA组合物及其使用方法
US9068184B2 (en) 2011-06-21 2015-06-30 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibition of expression of protein C (PROC) genes
US20140227293A1 (en) 2011-06-30 2014-08-14 Trustees Of Boston University Method for controlling tumor growth, angiogenesis and metastasis using immunoglobulin containing and proline rich receptor-1 (igpr-1)
WO2013001372A2 (en) 2011-06-30 2013-01-03 University Of Oslo Methods and compositions for inhibition of activation of regulatory t cells
HUE030501T2 (en) 2011-07-06 2017-05-29 Sykehuset Sorlandet Hf EGFR Targeted Therapy
WO2013006861A1 (en) 2011-07-07 2013-01-10 University Of Georgia Research Foundation, Inc. Sorghum grain shattering gene and uses thereof in altering seed dispersal
US8853181B2 (en) 2011-07-21 2014-10-07 Albert Einstein College Of Medicine Of Yeshiva University Fidgetin-like 2 as a target to enhance wound healing
US9120858B2 (en) 2011-07-22 2015-09-01 The Research Foundation Of State University Of New York Antibodies to the B12-transcobalamin receptor
DE102011118024A1 (de) 2011-08-01 2013-02-07 Technische Universität Dresden Inhibitor der Expression der Pro-Caspase 1
PL2753696T3 (pl) 2011-09-06 2018-06-29 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Rodzina miRNA-212/132 jako cel terapeutyczny
AU2012305714A1 (en) 2011-09-09 2014-03-27 Biomed Realty, L.P. Methods and compositions for controlling assembly of viral proteins
US9644241B2 (en) 2011-09-13 2017-05-09 Interpace Diagnostics, Llc Methods and compositions involving miR-135B for distinguishing pancreatic cancer from benign pancreatic disease
US9701623B2 (en) 2011-09-27 2017-07-11 Alnylam Pharmaceuticals, Inc. Di-aliphatic substituted pegylated lipids
WO2013049389A1 (en) 2011-09-27 2013-04-04 Yale University Compositions and methods for transient expression of recombinant rna
EP2768958B1 (en) 2011-10-18 2019-08-14 Dicerna Pharmaceuticals, Inc. Amine cationic lipids and uses thereof
EP2776565A1 (en) 2011-11-08 2014-09-17 Quark Pharmaceuticals, Inc. Methods and compositions for treating diseases, disorders or injury of the nervous system
US9006199B2 (en) 2011-11-14 2015-04-14 Silenseed Ltd. Methods and compositions for treating prostate cancer
ES2800065T3 (es) 2011-11-18 2020-12-23 Alnylam Pharmaceuticals Inc Agentes de iARN, composiciones y métodos de uso de los mismos para tratar enfermedades asociadas con transtiretina (TTR)
WO2013082529A1 (en) 2011-12-02 2013-06-06 Yale University Enzymatic synthesis of poly(amine-co-esters) and methods of use thereof for gene delivery
CA2858336A1 (en) 2012-01-01 2013-07-04 Qbi Enterprises Ltd. Endo180-targeted particles for selective delivery of therapeutic and diagnostic agents
US9464291B2 (en) * 2012-01-06 2016-10-11 University Of South Alabama Methods and compositions for the treatment of cancer
AU2013208720A1 (en) 2012-01-09 2014-07-24 Arrowhead Research Corporation RNAi agents to treat Beta-Catenin related diseases
AU2013208012A1 (en) 2012-01-12 2014-07-03 Quark Pharmaceuticals, Inc. Combination therapy for treating hearing and balance disorders
US20150126438A1 (en) 2012-01-24 2015-05-07 Beth Israel Deaconess Medical Center, Inc. Novel ChREBP Isoforms and Methods Using the Same
WO2013138463A1 (en) 2012-03-14 2013-09-19 University Of Central Florida Research Foundation, Inc. Neurofibromatoses therapeutic agents and screening for same
US20150031750A1 (en) 2012-03-15 2015-01-29 The Scripps Research Institute Treatment of brain derived neurotrophic factor (bdnf) related diseases by inhibition of natural antisense transcript to bdnf
WO2013158859A1 (en) 2012-04-18 2013-10-24 Cell Signaling Technology, Inc. Egfr and ros1 in cancer
US9980942B2 (en) 2012-05-02 2018-05-29 Children's Hospital Medical Center Rejuvenation of precursor cells
US20150299696A1 (en) 2012-05-02 2015-10-22 Sirna Therapeutics, Inc. SHORT INTERFERING NUCLEIC ACID (siNA) COMPOSITIONS
ES2716818T3 (es) 2012-05-22 2019-06-17 Olix Pharmaceuticals Inc Molécula de ácido nucleico inductora de interferencias de arn capaz de penetrar en las células y uso de la misma
US9726661B2 (en) 2012-07-06 2017-08-08 Institut Gustave-Roussy Simultaneous detection of cannibalism and senescence as prognostic marker for cancer
WO2014018375A1 (en) 2012-07-23 2014-01-30 Xenon Pharmaceuticals Inc. Cyp8b1 and uses thereof in therapeutic and diagnostic methods
JP6431480B2 (ja) * 2012-08-31 2018-11-28 ザ ジェネラル ホスピタル コーポレイション アルツハイマー病の治療および診断のためのビオチン複合体
GB201215857D0 (en) 2012-09-05 2012-10-24 Sylentis Sau siRNA and their use in methods and compositions for the treatment and/or prevention of eye conditions
SG11201501385UA (en) 2012-09-05 2015-03-30 Sylentis Sau Sirna and their use in methods and compositions for the treatment and/or prevention of eye conditions
ES2704855T3 (es) 2012-09-12 2019-03-20 Quark Pharmaceuticals Inc Moléculas de oligonucleótido de doble cadena para p53 y métodos de uso de las mismas
WO2014043289A2 (en) 2012-09-12 2014-03-20 Quark Pharmaceuticals, Inc. Double-stranded oligonucleotide molecules to ddit4 and methods of use thereof
SG10202011046RA (en) 2012-09-21 2020-12-30 Intensity Therapeutics Inc Method of treating cancer
EP3795694A3 (en) * 2012-10-02 2021-06-23 The General Hospital Corporation d/b/a Massachusetts General Hospital Methods relating to dna-sensing pathway related conditions
WO2014055825A1 (en) 2012-10-04 2014-04-10 The United States Of America, As Represented By The Secretary, Department Of Health & Human Services A formulation of mycobacterial components as an adjuvant for inducing th17 responses
WO2014068072A1 (en) 2012-10-31 2014-05-08 Institut Gustave-Roussy Identification, assessment and therapy of essential thrombocythemia with resistance to jak2 inhibitors
CA2930339A1 (en) * 2012-11-13 2014-05-22 Jan Lotvall Delivery of therapeutic agent
DE102012022596B4 (de) 2012-11-15 2017-05-04 Friedrich-Schiller-Universität Jena Neue zellspezifisch wirksame Nukleotid-Moleküle und Applikationskit zu deren Anwendung
PL2920201T3 (pl) 2012-11-15 2020-10-05 Apellis Pharmaceuticals, Inc. Długo działające analogi kompstatyny oraz powiązane kompozycje i sposoby
ES2884925T3 (es) 2012-11-27 2021-12-13 Childrens Medical Ct Corp Elementos reguladores distales de BCL11A como dianas para la reinducción de la hemoglobina fetal
US9970002B2 (en) 2012-12-12 2018-05-15 Massachusetts Institute Of Technology Compositions and methods for functional nucleic acid delivery
KR102228358B1 (ko) 2012-12-21 2021-03-16 쉬케후세트 솔란데트 에이치에프 신경 장애 및 통증의 egfr 표적 치료
US9206423B2 (en) * 2012-12-30 2015-12-08 The Regents Of The University Of California Methods of modulating compliance of the trabecular meshwork
US10258682B2 (en) 2013-01-16 2019-04-16 The United States Of America, As Represented By The Secretary, Department Of Health & Human Services Attenuated chlamydia vaccine
DE102013003869B4 (de) 2013-02-27 2016-11-24 Friedrich-Schiller-Universität Jena Verfahren zur gezielten Abtötung von Zellen durch zur mRNA-Anbindung ausgerichtete Nukleotid-Moleküle sowie Nukleotid-Moleküle und Applikationskit für solche Verwendung
CA3120574A1 (en) 2013-03-14 2014-09-25 Dicerna Pharmaceuticals, Inc. Process for formulating an anionic agent
WO2014152391A1 (en) 2013-03-15 2014-09-25 Apellis Pharmaceuticals, Inc. Cell-penetrating compstatin analogs and uses thereof
WO2014150751A2 (en) * 2013-03-15 2014-09-25 Novartis Ag Biomarkers associated with brm inhibition
KR20150132198A (ko) * 2013-03-15 2015-11-25 제넨테크, 인크. 브로모도메인-함유 단백질 brd7 및 brd9의 억제에 의한 th2-매개 질환의 치료
WO2014160871A2 (en) 2013-03-27 2014-10-02 The General Hospital Corporation Methods and agents for treating alzheimer's disease
US20160146806A1 (en) 2013-05-17 2016-05-26 Amplimmune, Inc. Receptors for b7-h4
AU2014302038B2 (en) 2013-06-25 2019-11-14 Epiaxis Therapeutics Pty Ltd Methods and compositions for modulating cancer stem cells
TW201534578A (zh) 2013-07-08 2015-09-16 Daiichi Sankyo Co Ltd 新穎脂質
UA122662C2 (uk) 2013-07-19 2020-12-28 Монсанто Текнолоджі Ллс Композиція та спосіб боротьби з leptinotarsa
WO2015013510A1 (en) 2013-07-25 2015-01-29 Ecole Polytechnique Federale De Lausanne Epfl High aspect ratio nanofibril materials
WO2015015496A1 (en) 2013-07-31 2015-02-05 Qbi Enterprises Ltd. Sphingolipid-polyalkylamine-oligonucleotide compounds
WO2015015498A1 (en) 2013-07-31 2015-02-05 Qbi Enterprises Ltd. Methods of use of sphingolipid polyalkylamine oligonucleotide compounds
BR112016004093A2 (pt) 2013-08-28 2017-10-17 Ionis Pharmaceuticals Inc modulação da expressão de pré-calicreína (pkk)
KR102435648B1 (ko) 2013-09-11 2022-08-25 이글 바이오로직스 인코퍼레이티드 점도저하제를 함유하는 액체 단백질 제형
CN105764513A (zh) 2013-09-18 2016-07-13 堪培拉大学 干细胞调控ii
KR102298475B1 (ko) 2013-10-04 2021-09-06 노파르티스 아게 RNA 간섭에 사용하기 위한 RNAi 작용제를 위한 3' 말단 캡
JP6546161B2 (ja) 2013-10-04 2019-07-17 ノバルティス アーゲー B型肝炎ウイルスを治療するための有機化合物
EP2865756A1 (en) 2013-10-22 2015-04-29 Sylentis, S.A.U. siRNA and their use in methods and compositions for inhibiting the expression of the FLAP gene.
EP2865758A1 (en) 2013-10-22 2015-04-29 Sylentis, S.A.U. siRNA and their use in methods and compositions for inhibiting the expression of the ORAI1 gene
EP2865757A1 (en) 2013-10-22 2015-04-29 Sylentis, S.A.U. siRNA and their use in methods and compositions for inhibiting the expression of the PDK1 gene.
EP3068407A1 (en) 2013-11-11 2016-09-21 Sirna Therapeutics, Inc. Systemic delivery of myostatin short interfering nucleic acids (sina) conjugated to a lipophilic moiety
US10441637B2 (en) 2013-11-21 2019-10-15 Sena Research, Inc. Methods for structural determination of selenium derivatized nucleic acid complexes
EP3077511A4 (en) 2013-12-06 2017-07-05 Dicerna Pharmaceuticals Inc. Methods and compositions for the specific inhibition of transthyretin (ttr) by double-stranded rna
CN104830906B (zh) 2014-02-12 2018-09-04 北京维通达生物技术有限公司 一种重编程获得功能性人肝脏实质细胞的方法
WO2015132303A1 (en) 2014-03-04 2015-09-11 Sylentis Sau Sirnas and their use in methods and compositions for the treatment and/or prevention of eye conditions
US20170016025A1 (en) 2014-03-11 2017-01-19 Cellectis Method for generating t-cells compatible for allogenic transplantation
DK3119887T3 (da) * 2014-03-20 2019-05-20 Oommen Varghese Forbedrede korte interfererende ribonukleinsyremolekyler
CA2946719C (en) 2014-03-25 2023-09-26 Arcturus Therapeutics, Inc. Una oligomers having reduced off-target effects in gene silencing
US9856475B2 (en) 2014-03-25 2018-01-02 Arcturus Therapeutics, Inc. Formulations for treating amyloidosis
JP6771387B2 (ja) 2014-03-25 2020-10-21 アークトゥラス・セラピューティクス・インコーポレイテッドArcturus Therapeutics,Inc. 遺伝子サイレンシング用トランスサイレチン対立遺伝子選択的unaオリゴマー
CN110506752B (zh) 2014-04-01 2022-02-18 孟山都技术公司 用于控制虫害的组合物和方法
PL3757214T3 (pl) 2014-04-01 2022-09-12 Biogen Ma Inc. Kompozycje do modulowania ekspresji sod-1
CN112852812A (zh) 2014-04-25 2021-05-28 儿童医疗中心有限公司 治疗血红蛋白病的组合物和方法
BR112016022855B1 (pt) 2014-05-01 2022-08-02 Ionis Pharmaceuticals, Inc Compostos e composições para modular a expressão de pkk e seus usos
WO2015168674A1 (en) 2014-05-02 2015-11-05 Research Institute At Nationwide Children's Hospital Compositions and methods for anti-lyst immunomodulation
WO2015175545A1 (en) 2014-05-12 2015-11-19 The Johns Hopkins University Highly stable biodegradable gene vector platforms for overcoming biological barriers
JP6763780B2 (ja) 2014-05-12 2020-09-30 ザ・ジョンズ・ホプキンス・ユニバーシティー 合成脳浸透遺伝子ベクターの操作
WO2015184105A1 (en) 2014-05-29 2015-12-03 Trustees Of Dartmouth College Method for selectively inhibiting acat1 in the treatment of neurodegenerative diseases
RU2754955C2 (ru) 2014-07-29 2021-09-08 Монсанто Текнолоджи Ллс Композиции и способы борьбы с насекомыми-вредителями
WO2016019126A1 (en) 2014-07-30 2016-02-04 The Research Foundation For The State University Of New York System and method for delivering genetic material or protein to cells
WO2016023974A1 (de) 2014-08-14 2016-02-18 Friedrich-Schiller-Universität Jena Peptid zur verwendung in der reduktion von nebenwirkungen in form von immunstimulatorischen reaktionen/effekten
CN107106517A (zh) 2014-08-25 2017-08-29 堪培拉大学 用于调节癌干细胞的组合物及其用途
US10060921B2 (en) 2014-08-29 2018-08-28 Alnylam Pharmaceuticals, Inc. Methods of treating transthyretin (TTR) mediated amyloidosis
JP6841753B2 (ja) 2014-09-15 2021-03-10 ザ チルドレンズ メディカル センター コーポレーション ヒストンh3−リジントリメチル化を除去することによって体細胞核移入(scnt)効率を増加させるための方法および組成物
US20170296561A1 (en) 2014-09-25 2017-10-19 Cold Spring Harbor Laboratory Treatment of rett syndrome
CA2962768C (en) 2014-10-01 2023-10-10 Alyssa M. Larson Polysaccharide and nucleic acid formulations containing viscosity-lowering agents
US20160101128A1 (en) 2014-10-10 2016-04-14 Idera Pharmaceuticals, Inc. Treatment of cancer using tlr9 agonist with checkpoint inhibitors
US20170304459A1 (en) 2014-10-10 2017-10-26 Alnylam Pharmaceuticals, Inc. Methods and compositions for inhalation delivery of conjugated oligonucleotide
JOP20200115A1 (ar) 2014-10-10 2017-06-16 Alnylam Pharmaceuticals Inc تركيبات وطرق لتثبيط التعبير الجيني عن hao1 (حمض أوكسيداز هيدروكسيلي 1 (أوكسيداز جليكولات))
WO2016060919A1 (en) 2014-10-14 2016-04-21 The Board Of Regents Of The University Of Texas System Allele selective inhibition of mutant c9orf72 foci expression by duplex rnas targeting the expanded hexanucleotide repeat
EP3209794A1 (en) 2014-10-22 2017-08-30 Katholieke Universiteit Leuven KU Leuven Research & Development Modulating adipose tissue and adipogenesis
JOP20200092A1 (ar) 2014-11-10 2017-06-16 Alnylam Pharmaceuticals Inc تركيبات iRNA لفيروس الكبد B (HBV) وطرق لاستخدامها
AU2015346281B2 (en) 2014-11-12 2021-12-02 Nmc, Inc. Transgenic plants with engineered redox sensitive modulation of photosynthetic antenna complex pigments and methods for making the same
WO2016081444A1 (en) 2014-11-17 2016-05-26 Alnylam Pharmaceuticals, Inc. Apolipoprotein c3 (apoc3) irna compositions and methods of use thereof
WO2016081621A1 (en) 2014-11-18 2016-05-26 Yale University Formulations for targeted release of agents under low ph conditions and methods of use thereof
CA2968531A1 (en) 2014-11-21 2016-05-26 Northwestern University The sequence-specific cellular uptake of spherical nucleic acid nanoparticle conjugates
WO2016094374A1 (en) * 2014-12-09 2016-06-16 The Board Of Regents Of The University Of Texas System Compositions and mentods for treatment of friedreich's ataxia
US10264976B2 (en) 2014-12-26 2019-04-23 The University Of Akron Biocompatible flavonoid compounds for organelle and cell imaging
US10792299B2 (en) 2014-12-26 2020-10-06 Nitto Denko Corporation Methods and compositions for treating malignant tumors associated with kras mutation
US20180002702A1 (en) * 2014-12-26 2018-01-04 Nitto Denko Corporation Methods and compositions for treating malignant tumors associated with kras mutation
RU2723049C2 (ru) 2015-01-22 2020-06-08 Монсанто Текнолоджи Ллс Композиции и способы борьбы с leptinotarsa
WO2016161299A1 (en) 2015-04-01 2016-10-06 Arcturus Therapeutics, Inc. Therapeutic una oligomers and uses thereof
WO2016168286A1 (en) 2015-04-13 2016-10-20 Alnylam Pharmaceuticals, Inc. Angiopoietin-like 3 (angptl3) irna compositions and methods of use thereof
WO2016168197A1 (en) 2015-04-15 2016-10-20 Yale University Compositions for enhancing delivery of agents across the blood brain barrier and methods of use thereof
WO2016179394A1 (en) 2015-05-05 2016-11-10 Malik Mohammad Tariq Anti-nucleolin agent-conjugated nanoparticles as radio-sensitizers and mri and/or x-ray contrast agents
EP3294873B1 (en) 2015-05-08 2020-08-19 The Children's Medical Center Corporation Targeting bcl11a enhancer functional regions for fetal hemoglobin reinduction
US10933049B2 (en) 2015-06-03 2021-03-02 The University Of Queensland Mobilizing agents and uses therefor
AU2016275046B2 (en) 2015-06-10 2022-07-28 Board Of Regents, The University Of Texas System Use of exosomes for the treatment of disease
JP6983066B2 (ja) 2015-06-30 2021-12-17 忠三 岸本 新規な肺疾患治療剤および/またはそのスクリーニング方法
EP4092119A3 (en) 2015-07-10 2023-03-22 Ionis Pharmaceuticals, Inc. Modulators of diacyglycerol acyltransferase 2 (dgat2)
WO2017015671A1 (en) 2015-07-23 2017-01-26 Arcturus Therapeutics, Inc. Compositions for treating amyloidosis
HUE051998T2 (hu) 2015-07-31 2021-04-28 Alnylam Pharmaceuticals Inc Transztiretin (TTR) iRNS készítmények és eljárások azok alkalmazása TTR-asszociált betegségek kezelésére vagy megelõzésére
CN108366966A (zh) 2015-08-24 2018-08-03 光环生物干扰疗法公司 用于调节基因表达的多核苷酸纳米颗粒及其用途
MA44908A (fr) 2015-09-08 2018-07-18 Sylentis Sau Molécules d'arnsi et leur utilisation dans des procédés et des compositions pour inhiber l'expression du gène nrarp
GB201516685D0 (en) * 2015-09-21 2015-11-04 Varghese Oommen P And Oommen Oommen P Nucleic acid molecules with enhanced activity
US10086063B2 (en) 2015-09-23 2018-10-02 Regents Of The University Of Minnesota Methods of making and using live attenuated viruses
CN108601823A (zh) 2015-09-23 2018-09-28 麻省理工学院 用于改性树枝状聚合物纳米颗粒疫苗递送的组合物和方法
US10383935B2 (en) 2015-09-23 2019-08-20 Regents Of The University Of Minnesota Methods of making and using live attenuated viruses
JP2018535655A (ja) 2015-09-29 2018-12-06 アムジエン・インコーポレーテツド Asgr阻害剤
JOP20210043A1 (ar) * 2015-10-01 2017-06-16 Arrowhead Pharmaceuticals Inc تراكيب وأساليب لتثبيط تعبير جيني للـ lpa
KR20180090785A (ko) 2015-10-07 2018-08-13 아펠리스 파마슈티컬스 인코포레이티드 투여 요법
WO2017085550A1 (en) 2015-11-16 2017-05-26 Olix Pharmaceuticals, Inc. Treatment of age-related macular degeneration using rna complexes that target myd88 or tlr3
WO2017095751A1 (en) 2015-12-02 2017-06-08 Partikula Llc Compositions and methods for modulating cancer cell metabolism
EP3386544B1 (en) 2015-12-10 2020-11-25 Fibrogen, Inc. Methods for treatment of motor neuron diseases
US20180344638A1 (en) 2015-12-18 2018-12-06 Samyang Biopharmaceuticals Corporation Method for preparing polymeric micelle containing anionic drug
BR102017001164A2 (pt) 2016-01-26 2019-03-06 Embrapa - Empresa Brasileira De Pesquisa Agropecuária Composições de rna de fita dupla para controle de diaphorina citri e métodos de uso.
JP2019503394A (ja) 2016-01-31 2019-02-07 ユニバーシティ・オブ・マサチューセッツUniversity Of Massachusetts 分岐オリゴヌクレオチド
JP7003044B2 (ja) 2016-02-02 2022-01-20 オリックス ファーマシューティカルズ,インコーポレーテッド Angpt2およびpdgfbを標的化するrna複合体を用いる血管新生関連疾患の処置
EP3411480A4 (en) 2016-02-02 2020-01-22 Olix Pharmaceuticals, Inc. TREATMENT OF ATOPIC DERMATITIS AND ASTHMA USING RNA COMPLEXES THAT TARGETE IL4R, TRPA1, OR F2RL1
US20170360815A1 (en) 2016-02-25 2017-12-21 Applied Biological Laboratories, Inc. Compositions and methods for protecting against airborne pathogens and irritants
US20170246262A1 (en) 2016-02-25 2017-08-31 Applied Biological Laboratories, Inc. Compositions and methods for protecting against airborne pathogens and irritants
JP2019512014A (ja) 2016-02-26 2019-05-09 イェール ユニバーシティーYale University がん診断法および治療法におけるpiRNAを使用する組成物および方法
WO2017151623A1 (en) 2016-03-01 2017-09-08 Alexion Pharmaceuticals, Inc. Biodegradable activated polymers for therapeutic delivery
EP3423568A4 (en) 2016-03-04 2019-11-13 University Of Louisville Research Foundation, Inc. METHOD AND COMPOSITIONS FOR EX-VIVO REPRODUCTION OF VERY SMALL EMBRYONAL STEM CELLS (VSELS)
TWI783434B (zh) 2016-03-07 2022-11-11 美商愛羅海德製藥公司 治療性化合物之標靶性配體
BR112018068461A2 (pt) 2016-03-15 2019-01-22 Mersana Therapeutics Inc conjugado, composição farmacêutica, métodos para preparação de um conjugado e para alívio de um sintoma de um câncer.
MA45470A (fr) 2016-04-01 2019-02-06 Avidity Biosciences Llc Acides nucléiques kras et leurs utilisations
MA45328A (fr) 2016-04-01 2019-02-06 Avidity Biosciences Llc Compositions acide nucléique-polypeptide et utilisations de celles-ci
WO2017173453A1 (en) 2016-04-01 2017-10-05 The Brigham And Women's Hospital, Inc. Stimuli-responsive nanoparticles for biomedical applications
MA45349A (fr) 2016-04-01 2019-02-06 Avidity Biosciences Llc Acides nucléiques egfr et leurs utilisations
MA45469A (fr) 2016-04-01 2019-02-06 Avidity Biosciences Llc Acides nucléiques de bêta-caténine et leurs utilisations
DK3440090T3 (da) 2016-04-06 2022-12-19 Ohio State Innovation Foundation RNA-ligand-præsenterende exosomere til specifik levering af terapeutika til en celle ved RNA-nanoteknologi
US10301628B2 (en) 2016-04-11 2019-05-28 Olix Pharmaceuticals, Inc. Treatment of idiopathic pulmonary fibrosis using RNA complexes that target connective tissue growth factor
US11234994B2 (en) 2016-04-14 2022-02-01 Benitec Biopharma Limited Reagents for treatment of oculopharyngeal muscular dystrophy (OPMD) and use thereof
WO2017189870A1 (en) 2016-04-27 2017-11-02 Massachusetts Institute Of Technology Stable nanoscale nucleic acid assemblies and methods thereof
WO2017197128A1 (en) 2016-05-11 2017-11-16 Yale University Poly(amine-co-ester) nanoparticles and methods of use thereof
KR101916652B1 (ko) 2016-06-29 2018-11-08 올릭스 주식회사 작은 간섭 rna의 rna 간섭효과 증진용 화합물 및 이의 용도
WO2018006005A1 (en) 2016-06-30 2018-01-04 Oncorus, Inc. Pseudotyped oncolytic viral delivery of therapeutic polypeptides
RU2627179C1 (ru) * 2016-07-28 2017-08-03 федеральное государственное бюджетное учреждение "Федеральный научно-исследовательский центр эпидемиологии и микробиологии имени почетного академика Н.Ф. Гамалеи" Министерства здравоохранения Российской Федерации ТЕСТ-СИСТЕМА ДЛЯ ОПРЕДЕЛЕНИЯ РНК ИНТЕРФЕРОНА λ, ИНТЕРЛЕЙКИНА IL23 И ПРОТИВОВИРУСНОГО БЕЛКА MxA
WO2018020012A1 (en) 2016-07-29 2018-02-01 Danmarks Tekniske Universitet Methods for decoupling cell growth from production of biochemicals and recombinant polypeptides
CN110087665A (zh) 2016-08-03 2019-08-02 H·李·莫菲特癌症中心与研究所公司 Tlr9靶向治疗
US11364304B2 (en) 2016-08-25 2022-06-21 Northwestern University Crosslinked micellar spherical nucleic acids
PT3506909T (pt) 2016-09-02 2022-08-16 Dicerna Pharmaceuticals Inc Análogos de 4¿-fosfato e oligonucleótidos compreendendo os mesmos
JP6989521B2 (ja) 2016-09-02 2022-01-05 アローヘッド ファーマシューティカルズ インコーポレイテッド 標的化リガンド
US10933081B2 (en) 2016-09-21 2021-03-02 Alnylam Pharmaceuticals, Inc. Myostatin iRNA compositions and methods of use thereof
US11260134B2 (en) 2016-09-29 2022-03-01 National University Corporation Tokyo Medical And Dental University Double-stranded nucleic acid complex having overhang
WO2018098352A2 (en) 2016-11-22 2018-05-31 Jun Oishi Targeting kras induced immune checkpoint expression
US11135307B2 (en) 2016-11-23 2021-10-05 Mersana Therapeutics, Inc. Peptide-containing linkers for antibody-drug conjugates
WO2018107096A1 (en) 2016-12-08 2018-06-14 University Of Utah Research Foundation Staufen1 agents and associated methods
WO2018112470A1 (en) 2016-12-16 2018-06-21 The Brigham And Women's Hospital, Inc. Co-delivery of nucleic acids for simultaneous suppression and expression of target genes
MX2019008252A (es) 2017-01-10 2019-09-06 Arrowhead Pharmaceuticals Inc Agentes de interferencia de acido ribonucleico (iarn) de alfa-1 antitripsina (aat), composiciones que incluyen agentes de iarn aat y metodos de uso.
EP3580339A4 (en) * 2017-02-10 2020-12-23 Research & Business Foundation Sungkyunkwan University LONG DOUBLE STRAND RNA FOR RNA INTERFERENCE
DE102017103383A1 (de) 2017-02-20 2018-08-23 aReNA-Bio GbR (vertretungsberechtigter Gesellschafter: Dr. Heribert Bohlen, 50733 Köln) System und Verfahren zur Zelltyp-spezifischen Translation von RNA-Molekülen in Eukaryoten
US11613754B2 (en) * 2017-02-20 2023-03-28 Northwestern University Toxic RNAi active seed sequences for killing cancer cells
TW201834697A (zh) 2017-02-28 2018-10-01 美商梅爾莎納醫療公司 Her2標靶抗體-藥物結合物之組合療法
US11261441B2 (en) 2017-03-29 2022-03-01 Bluebird Bio, Inc. Vectors and compositions for treating hemoglobinopathies
KR20190139931A (ko) 2017-04-07 2019-12-18 아펠리스 파마슈티컬스 인코포레이티드 투여 요법 및 관련 조성물 및 방법
CN110945128B (zh) 2017-04-14 2023-11-03 代表亚利桑那大学的亚利桑那董事会 用于治疗肺纤维化的组合物和方法
US11324820B2 (en) 2017-04-18 2022-05-10 Alnylam Pharmaceuticals, Inc. Methods for the treatment of subjects having a hepatitis b virus (HBV) infection
US11433131B2 (en) 2017-05-11 2022-09-06 Northwestern University Adoptive cell therapy using spherical nucleic acids (SNAs)
US11788087B2 (en) 2017-05-25 2023-10-17 The Children's Medical Center Corporation BCL11A guide delivery
US10844377B2 (en) * 2017-06-23 2020-11-24 University Of Massachusetts Two-tailed self-delivering siRNA
TN2019000308A1 (en) 2017-07-06 2021-05-07 Arrowhead Pharmaceuticals Inc RNAi AGENTS FOR INHIBITING EXPRESSION OF ALPHA-ENaC AND METHODS OF USE
AU2018301829A1 (en) 2017-07-13 2020-02-27 Alnylam Pharmaceuticals, Inc. Methods for inhibition of HAO1 (hydroxyacid oxidase 1 (glycolate oxidase) gene expression
US11104700B2 (en) 2017-07-17 2021-08-31 Oxford University Innovation Limited Oligonucleotides
US11110114B2 (en) 2017-07-17 2021-09-07 Oxford University Innovation Limited Dinucleotides
JP7438103B2 (ja) 2017-09-11 2024-02-26 アローヘッド ファーマシューティカルズ インコーポレイテッド アポリポタンパク質C-III(APOC3)の発現を阻害するためのRNAi剤および組成物
WO2019060442A1 (en) 2017-09-19 2019-03-28 Alnylam Pharmaceuticals, Inc. COMPOSITIONS AND METHODS FOR TREATMENT OF TRANSTHYRETIN MEDIATED AMYLOSIS (TTR)
WO2019068326A1 (en) 2017-10-05 2019-04-11 Université D'aix-Marseille INHIBITORS OF LSD1 FOR THE TREATMENT AND PREVENTION OF CARDIOMYOPATHIES
CR20200163A (es) 2017-10-20 2020-11-02 Dicerna Pharmaceuticals Inc Oligonucleótidos y composiciones para tratar la infección por hepatitis
WO2019100053A1 (en) 2017-11-20 2019-05-23 University Of Georgia Research Foundation, Inc. Compositions and methods for modulating hif-2α to improve muscle generation and repair
WO2019104289A1 (en) 2017-11-27 2019-05-31 Mersana Therapeutics, Inc. Pyrrolobenzodiazepine antibody conjugates
US11198869B2 (en) 2017-12-01 2021-12-14 The Texas A&M University System Angelman syndrome antisense treatment
KR102623786B1 (ko) 2017-12-06 2024-01-11 어비디티 바이오사이언시스 인크. 근위축증 및 근긴장성 이영양증을 치료하는 조성물 및 방법
CA3086485A1 (en) 2017-12-21 2019-06-27 Alnylam Pharmaceuticals, Inc. Chirally-enriched double-stranded rna agents
CN111757757A (zh) 2017-12-21 2020-10-09 梅尔莎纳医疗公司 吡咯并苯并二氮呯抗体共轭物
US10960086B2 (en) 2017-12-28 2021-03-30 Augusta University Research Institute, Inc. Aptamer compositions and methods of use thereof
EP3731850A4 (en) 2017-12-29 2021-12-01 Oncorus, Inc. ONCOLYTIC VIRUS DELIVERY OF THERAPEUTIC POLYPEPTIDES
EP3697423A4 (en) 2018-01-05 2021-08-11 Dicerna Pharmaceuticals, Inc. REDUCED EXPRESSION OF BETA-CATENIN AND IDO TO POTENTIALIZE IMMUNOTHERAPY
WO2019143621A1 (en) 2018-01-16 2019-07-25 Dicerna Pharmaceuticals, Inc. Compositions and methods for inhibiting aldh2 expression
CR20200346A (es) 2018-02-09 2020-10-19 Genentech Inc Oligonucleótidos para modular la expresión de tmem106b
WO2019215067A1 (en) 2018-05-07 2019-11-14 Roche Innovation Center Copenhagen A/S Massively parallel discovery methods for oligonucleotide therapeutics
CA3100995A1 (en) 2018-05-10 2019-11-14 The University Of Manchester Methods for assessing macular degeneration
JP2021527649A (ja) * 2018-06-14 2021-10-14 ユニバーシティー オブ ユタ リサーチ ファウンデーションUniversity of Utah Research Foundation Staufen1調節剤および関連する方法
AU2019321375A1 (en) 2018-08-13 2021-03-11 Alnylam Pharmaceuticals, Inc. Hepatitis B virus (HBV) dsRNA agent compositions and methods of use thereof
US20210205264A1 (en) 2018-09-04 2021-07-08 H. Lee Moffitt Cancer Center & Research Institute Inc. Use of delta-tocotrienol for treating cancer
EP3847650A1 (en) 2018-09-06 2021-07-14 The Broad Institute, Inc. Nucleic acid assemblies for use in targeted delivery
CA3117050A1 (en) 2018-10-29 2020-05-07 Mersana Therapeutics, Inc. Cysteine engineered antibody-drug conjugates with peptide-containing linkers
WO2020167593A1 (en) 2019-02-12 2020-08-20 Dicerna Pharmaceuticals, Inc. Methods and compositions for inhibiting expression of cyp27a1
CA3134486A1 (en) 2019-03-29 2020-10-08 Dicerna Pharmaceuticals, Inc. Compositions and methods for the treatment of kras associated diseases or disorders
US20220170025A1 (en) 2019-04-04 2022-06-02 Dicerna Pharmaceuticals Inc. Compositions and methods for inhibiting gene expression in the central nervous system
US11814464B2 (en) 2019-04-29 2023-11-14 Yale University Poly(amine-co-ester) polymers and polyplexes with modified end groups and methods of use thereof
CN113795581A (zh) 2019-05-03 2021-12-14 迪克纳制药公司 具有缩短的有义链的双链核酸抑制剂分子
US20200369759A1 (en) 2019-05-23 2020-11-26 Fibrogen, Inc. Methods of treatment of muscular dystrophies
US20220257790A1 (en) 2019-06-26 2022-08-18 Biorchestra Co., Ltd. Micellar nanoparticles and uses thereof
AU2020336992A1 (en) 2019-08-30 2022-04-14 Yale University Compositions and methods for delivery of nucleic acids to cells
WO2021049504A1 (ja) 2019-09-10 2021-03-18 第一三共株式会社 肝臓送達用GalNAc-オリゴヌクレオチドコンジュゲートおよび製造方法
IL291841A (en) 2019-10-02 2022-06-01 Dicerna Pharmaceuticals Inc Chemical modifications of tiny interfering rna with minimal fluoride content
US11017851B1 (en) 2019-11-26 2021-05-25 Cypress Semiconductor Corporation Silicon-oxide-nitride-oxide-silicon based multi level non-volatile memory device and methods of operation thereof
CA3163646A1 (en) 2019-12-24 2021-07-01 F. Hoffman-La Roche Ag Pharmaceutical combination of antiviral agents targeting hbv and/or an immune modulator for treatment of hbv
MX2022007908A (es) 2019-12-24 2022-07-21 Hoffmann La Roche Combinacion farmaceutica de un oligonucleotido terapeutico que actua sobre hbv y un agonista de tlr7 para el tratamiento de hbv.
JP2023511274A (ja) 2020-01-14 2023-03-17 シンセカイン インコーポレイテッド Il2オルソログおよび使用法
WO2021150300A1 (en) 2020-01-22 2021-07-29 Massachusetts Institute Of Technology Inducible tissue constructs and uses thereof
US11642407B2 (en) 2020-02-28 2023-05-09 Massachusetts Institute Of Technology Identification of variable influenza residues and uses thereof
CN116096889A (zh) 2020-03-18 2023-05-09 迪克纳制药公司 用于抑制angptl3表达的组合物和方法
EP4121063A1 (en) 2020-03-19 2023-01-25 Avidity Biosciences, Inc. Compositions and methods of treating facioscapulohumeral muscular dystrophy
EP4126066A1 (en) 2020-03-27 2023-02-08 Avidity Biosciences, Inc. Compositions and methods of treating muscle dystrophy
BR112022021020A2 (pt) 2020-04-22 2023-02-14 Iovance Biotherapeutics Inc Método de fabricação de um produto de terapia celular, métodos de tratamento do paciente com o produto de terapia celular de expansão e fabricado, e, método de fabricação de um produto de terapia celular
WO2021255262A1 (en) 2020-06-19 2021-12-23 Sylentis Sau siRNA AND COMPOSITIONS FOR PROPHYLACTIC AND THERAPEUTIC TREATMENT OF VIRUS DISEASES
KR20230042018A (ko) 2020-06-19 2023-03-27 예일 유니버시티 변형된 말단기를 갖는 폴리(아민-코-에스테르) 폴리머 및 향상된 폐 운반
US20220031633A1 (en) 2020-07-28 2022-02-03 Yale University Poly(amine-co-ester) polymeric particles for selective pulmonary delivery
TW202221120A (zh) 2020-08-04 2022-06-01 美商黛瑟納製藥公司 用於治療代謝症候群之組成物及方法
KR20230061389A (ko) 2020-08-04 2023-05-08 다이서나 파마수이티컬, 인크. 올리고뉴클레오티드의 전신 전달
MX2023001450A (es) 2020-08-04 2023-04-14 Dicerna Pharmaceuticals Inc Composiciones y metodos para la inhibicion de la expresion de plp1.
KR20230043877A (ko) 2020-08-05 2023-03-31 에프. 호프만-라 로슈 아게 B형 간염 환자의 올리고뉴클레오티드 치료
CN116323943A (zh) 2020-08-05 2023-06-23 迪克纳制药公司 用于抑制lpa表达的组合物和方法
CA3193424A1 (en) 2020-08-31 2022-03-03 Yale University Compositions and methods for delivery of nucleic acids to cells
EP3964204A1 (en) 2020-09-08 2022-03-09 Université d'Aix-Marseille Lsd1 inhibitors for use in the treatment and prevention of fibrosis of tissues
WO2022058447A1 (en) 2020-09-16 2022-03-24 The University Of Manchester Complementome assay
IL303195A (en) 2020-11-25 2023-07-01 Akagera Medicines Inc Lipid nanoparticles for delivery of nucleic acids and related methods of use
CN117295753A (zh) 2020-12-04 2023-12-26 基那奥生物公司 用于将核酸递送到细胞的组合物和方法
EP4015634A1 (en) 2020-12-15 2022-06-22 Sylentis, S.A.U. Sirna and compositions for prophylactic and therapeutic treatment of virus diseases
IL305414A (en) 2021-03-04 2023-10-01 Alnylam Pharmaceuticals Inc Angiopoietin-like 3 (ANGPTL3) IRNA compositions and methods of using them
CN117202894A (zh) 2021-03-31 2023-12-08 胭脂红治疗私人有限公司 负载至少两个不同核酸的细胞外囊泡
US20220340909A1 (en) 2021-04-12 2022-10-27 Boehringer Ingelheim International Gmbh Compositions and methods for inhibiting ketohexokinase (khk)
BR112023021109A2 (pt) 2021-04-14 2023-12-12 Dicerna Pharmaceuticals Inc Composições e métodos para modular a expressão de pnpla3
AU2022261359A1 (en) 2021-04-19 2023-10-12 Dicerna Pharmaceuticals, Inc. Compositions and methods for inhibiting nuclear receptor subfamily 1 group h member 3 (nr1h3) expression
KR20230130609A (ko) 2021-05-28 2023-09-12 노보 노르디스크 에이/에스 미토콘드리아 아미독심 환원 성분 1(marc1) 발현을 억제하기 위한 조성물 및 방법
CA3221923A1 (en) 2021-05-29 2022-12-08 1Globe Health Institute Llc Short duplex dna as a novel gene silencing technology and use thereof
AU2022285661A1 (en) 2021-05-29 2023-12-21 1Globe Health Institute Llc Asymmetric short duplex dna as a novel gene silencing technology and use thereof
AU2022299169A1 (en) 2021-06-23 2024-02-08 Beth Israel Deaconess Medical Center, Inc. Optimized anti-flt1 oligonucleotide compounds for treatment of preeclampsia and other angiogenic disorders
CA3229305A1 (en) 2021-08-16 2023-02-23 Vib Vzw Oligonucleotides for modulating synaptogyrin-3 expression
AR124636A1 (es) 2021-08-25 2023-04-19 Dicerna Pharmaceuticals Inc COMPOSICIONES Y MÉTODOS PARA INHIBIR LA EXPRESIÓN DE a-1 ANTITRIPSINA
WO2023049743A1 (en) 2021-09-21 2023-03-30 The Johns Hopkins University Dendrimer conjugates of small molecule biologics for intracellular delivery
WO2023083906A2 (en) 2021-11-11 2023-05-19 F. Hoffmann-La Roche Ag Pharmaceutical combinations for treatment of hbv
WO2023092089A1 (en) 2021-11-19 2023-05-25 Kist (Korea Institute Of Science And Technology) Therapeutic compounds for red blood cell-mediated delivery of an active pharmaceutical ingredient to a target cell
US20230272393A1 (en) 2021-12-01 2023-08-31 Dicerna Pharmaceuticals, Inc. Compositions and methods for modulating apoc3 expression
WO2023118546A2 (en) 2021-12-23 2023-06-29 Boehringer Ingelheim International Gmbh METHODS AND MOLECULES FOR RNA INTERFERENCE (RNAi)
WO2023159189A1 (en) 2022-02-18 2023-08-24 Yale University Branched poly(amine-co-ester) polymers for more efficient nucleic expression
GB202203627D0 (en) 2022-03-16 2022-04-27 Univ Manchester Agents for treating complement-related disorders
WO2023192872A1 (en) 2022-03-28 2023-10-05 Massachusetts Institute Of Technology Rna scaffolded wireframe origami and methods thereof
GB202204884D0 (en) 2022-04-04 2022-05-18 Fondo Ricerca Medica S R I Sirna targeting kcna1
WO2023201043A1 (en) 2022-04-15 2023-10-19 Dicerna Pharmaceuticals, Inc. Compositions and methods for modulating scap activity
US20230416742A1 (en) 2022-05-12 2023-12-28 Dicerna Phrmaceuticals, Inc. Compositions and methods for inhibiting mapt expression
US20230416743A1 (en) 2022-05-13 2023-12-28 Dicerna Pharmaceuticals, Inc. Compositions and methods for inhibiting snca expression
US20240002858A1 (en) 2022-06-24 2024-01-04 Novo Nordisk A/S Compositions and methods for inhibiting transmembrane serine protease 6 (tmprss6) expression
WO2024040041A1 (en) 2022-08-15 2024-02-22 Dicerna Pharmaceuticals, Inc. Regulation of activity of rnai molecules

Family Cites Families (136)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2003006A (en) * 1933-04-11 1935-05-28 Michelson Barnett Samuel Water tank cover
US4469863A (en) * 1980-11-12 1984-09-04 Ts O Paul O P Nonionic nucleic acid alkyl and aryl phosphonates and processes for manufacture and use thereof
US5208149A (en) * 1983-10-20 1993-05-04 The Research Foundation Of State University Of New York Nucleic acid constructs containing stable stem and loop structures
GB8704365D0 (en) * 1987-02-25 1987-04-01 Exxon Chemical Patents Inc Zeolite l preparation
US5712257A (en) * 1987-08-12 1998-01-27 Hem Research, Inc. Topically active compositions of mismatched dsRNAs
IE66830B1 (en) 1987-08-12 1996-02-07 Hem Res Inc Topically active compositions of double-stranded RNAs
US5703055A (en) * 1989-03-21 1997-12-30 Wisconsin Alumni Research Foundation Generation of antibodies through lipid mediated DNA delivery
DK0497875T3 (da) * 1989-10-24 2000-07-03 Gilead Sciences Inc 2'-modificerede oligonukleotider
US5457189A (en) * 1989-12-04 1995-10-10 Isis Pharmaceuticals Antisense oligonucleotide inhibition of papillomavirus
EP0604409B1 (en) 1990-01-11 2004-07-14 Isis Pharmaceuticals, Inc. Oligonucleotide analogs for detecting and modulating rna activity and gene expression
US5670633A (en) * 1990-01-11 1997-09-23 Isis Pharmaceuticals, Inc. Sugar modified oligonucleotides that detect and modulate gene expression
US5514577A (en) * 1990-02-26 1996-05-07 Isis Pharmaceuticals, Inc. Oligonucleotide therapies for modulating the effects of herpes viruses
EP0552178B1 (en) * 1990-10-12 1997-01-02 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Modified ribozymes
FR2675803B1 (fr) 1991-04-25 1996-09-06 Genset Sa Oligonucleotides fermes, antisens et sens et leurs applications.
WO1994008003A1 (en) * 1991-06-14 1994-04-14 Isis Pharmaceuticals, Inc. ANTISENSE OLIGONUCLEOTIDE INHIBITION OF THE ras GENE
FR2685346B1 (fr) * 1991-12-18 1994-02-11 Cis Bio International Procede de preparation d'arn double-brin, et ses applications.
WO1993018052A1 (en) 1992-03-05 1993-09-16 Isis Pharmaceuticals, Inc. Covalently cross-linked oligonucleotides
US5792751A (en) * 1992-04-13 1998-08-11 Baylor College Of Medicine Tranformation of cells associated with fluid spaces
US20040054156A1 (en) * 1992-05-14 2004-03-18 Kenneth Draper Method and reagent for inhibiting hepatitis B viral replication
US20030068301A1 (en) * 1992-05-14 2003-04-10 Kenneth Draper Method and reagent for inhibiting hepatitis B virus replication
US20030206887A1 (en) * 1992-05-14 2003-11-06 David Morrissey RNA interference mediated inhibition of hepatitis B virus (HBV) using short interfering nucleic acid (siNA)
US5693535A (en) * 1992-05-14 1997-12-02 Ribozyme Pharmaceuticals, Inc. HIV targeted ribozymes
US20030171311A1 (en) * 1998-04-27 2003-09-11 Lawrence Blatt Enzymatic nucleic acid treatment of diseases or conditions related to hepatitis C virus infection
ATE171210T1 (de) 1992-07-02 1998-10-15 Hybridon Inc Selbststabilisierte oligonukleotide als therapeutika
US5652355A (en) 1992-07-23 1997-07-29 Worcester Foundation For Experimental Biology Hybrid oligonucleotide phosphorothioates
AU6080294A (en) 1992-12-31 1994-08-15 Texas Biotechnology Corporation Antisense molecules directed against genes of the (raf) oncogene family
US6056704A (en) 1993-03-03 2000-05-02 Ide; Masatake Foot-pressure massage stand
EP0616026A1 (en) 1993-03-19 1994-09-21 The Procter & Gamble Company Concentrated cleaning compositions
WO1995000638A2 (en) * 1993-06-23 1995-01-05 Genesys Pharma Inc. Antisense oligonucleotides and therapeutic use thereof in human immunodeficiency virus infection
FR2710074B1 (fr) 1993-09-15 1995-12-08 Rhone Poulenc Rorer Sa Gène GRB3-3, ses variants et leurs utilisations.
US5624803A (en) * 1993-10-14 1997-04-29 The Regents Of The University Of California In vivo oligonucleotide generator, and methods of testing the binding affinity of triplex forming oligonucleotides derived therefrom
US5801154A (en) * 1993-10-18 1998-09-01 Isis Pharmaceuticals, Inc. Antisense oligonucleotide modulation of multidrug resistance-associated protein
CA2176259A1 (en) 1993-11-16 1995-05-26 Lyle J. Arnold, Jr. Chimeric oligonucleoside compounds
US5908779A (en) * 1993-12-01 1999-06-01 University Of Connecticut Targeted RNA degradation using nuclear antisense RNA
US5578716A (en) 1993-12-01 1996-11-26 Mcgill University DNA methyltransferase antisense oligonucleotides
WO1995030746A1 (en) * 1994-05-10 1995-11-16 The General Hospital Corporation Antisense inhibition of hepatitis c virus
US6057153A (en) * 1995-01-13 2000-05-02 Yale University Stabilized external guide sequences
US5674683A (en) 1995-03-21 1997-10-07 Research Corporation Technologies, Inc. Stem-loop and circular oligonucleotides and method of using
US5624808A (en) * 1995-03-28 1997-04-29 Becton Dickinson And Company Method for identifying cells committed to apoptosis by determining cellular phosphotyrosine content
EP1489184A1 (en) 1995-06-07 2004-12-22 Inex Pharmaceutical Corp. Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer
EP0851919A1 (en) * 1995-09-20 1998-07-08 University of Massachusetts Worcester Antisense oligonucleotide chemotherapy for benign hyperplasia or cancer of the prostate
US5998203A (en) * 1996-04-16 1999-12-07 Ribozyme Pharmaceuticals, Inc. Enzymatic nucleic acids containing 5'-and/or 3'-cap structures
CN1214688A (zh) 1996-02-14 1999-04-21 伊希斯药物有限公司 糖修饰的缺口寡核苷酸
WO1997039120A2 (en) 1996-04-17 1997-10-23 Aronex Pharmaceuticals, Inc. Antisense inhibitors of vascular endothelial growth factor (vefg/vpf) expression
DE19618797C2 (de) 1996-05-10 2000-03-23 Bertling Wolf Vehikel zum Transport molekularer Substanz
US5898031A (en) * 1996-06-06 1999-04-27 Isis Pharmaceuticals, Inc. Oligoribonucleotides for cleaving RNA
US20040266706A1 (en) 2002-11-05 2004-12-30 Muthiah Manoharan Cross-linked oligomeric compounds and their use in gene modulation
DE19631919C2 (de) 1996-08-07 1998-07-16 Deutsches Krebsforsch Anti-Sinn-RNA mit Sekundärstruktur
US6225290B1 (en) * 1996-09-19 2001-05-01 The Regents Of The University Of California Systemic gene therapy by intestinal cell transformation
ATE329015T1 (de) * 1996-10-04 2006-06-15 Derek Nigel John Hart Enzyme mit s-adenosyl-l-homocystein-hydrolase- ähnlicher aktivität.
US5814500A (en) * 1996-10-31 1998-09-29 The Johns Hopkins University School Of Medicine Delivery construct for antisense nucleic acids and methods of use
EP0951536B1 (en) 1996-12-12 2007-01-24 Yissum Research Development Company Of The Hebrew University Of Jerusalem Synthetic antisense oligodeoxynucleotides and pharmaceutical compositions containing them
US20030064945A1 (en) * 1997-01-31 2003-04-03 Saghir Akhtar Enzymatic nucleic acid treatment of diseases or conditions related to levels of epidermal growth factor receptors
GB9703146D0 (en) * 1997-02-14 1997-04-02 Innes John Centre Innov Ltd Methods and means for gene silencing in transgenic plants
US6218142B1 (en) * 1997-03-05 2001-04-17 Michael Wassenegger Nucleic acid molecules encoding polypeptides having the enzymatic activity of an RNA-directed RNA polymerase (RDRP)
GB9710475D0 (en) 1997-05-21 1997-07-16 Zeneca Ltd Gene silencing
JP4236812B2 (ja) 1997-09-12 2009-03-11 エクシコン エ/エス オリゴヌクレオチド類似体
JP2002508299A (ja) 1997-09-19 2002-03-19 セクイター, インク. センスmRNA治療
GB9720148D0 (en) * 1997-09-22 1997-11-26 Innes John Centre Innov Ltd Gene silencing materials and methods
US6506559B1 (en) 1997-12-23 2003-01-14 Carnegie Institute Of Washington Genetic inhibition by double-stranded RNA
US6475726B1 (en) * 1998-01-09 2002-11-05 Cubist Pharmaceuticals, Inc. Method for identifying validated target and assay combinations for drug development
AUPP249298A0 (en) * 1998-03-20 1998-04-23 Ag-Gene Australia Limited Synthetic genes and genetic constructs comprising same I
WO1999049029A1 (en) 1998-03-20 1999-09-30 Benitec Australia Ltd Control of gene expression
US20040214330A1 (en) * 1999-04-07 2004-10-28 Waterhouse Peter Michael Methods and means for obtaining modified phenotypes
EP1068311B2 (en) 1998-04-08 2020-12-09 Commonwealth Scientific and Industrial Research Organisation Methods and means for obtaining modified phenotypes
EP1071753A2 (en) 1998-04-20 2001-01-31 Ribozyme Pharmaceuticals, Inc. Nucleic acid molecules with novel chemical compositions capable of modulating gene expression
AR020078A1 (es) 1998-05-26 2002-04-10 Syngenta Participations Ag Metodo para alterar la expresion de un gen objetivo en una celula de planta
GB9827152D0 (en) 1998-07-03 1999-02-03 Devgen Nv Characterisation of gene function using double stranded rna inhibition
AU1408300A (en) 1998-11-24 2000-06-13 Hisamitsu Pharmaceutical Co. Inc. Hiv infection inhibitors
WO2000032619A1 (en) 1998-11-30 2000-06-08 Ribogene, Inc. Methods and compositions for identification of inhibitors of ribosome assembly
US6939712B1 (en) 1998-12-29 2005-09-06 Impedagen, Llc Muting gene activity using a transgenic nucleic acid
CA2361201A1 (en) 1999-01-28 2000-08-03 Medical College Of Georgia Research Institute, Inc. Composition and method for in vivo and in vitro attenuation of gene expression using double stranded rna
DE19956568A1 (de) 1999-01-30 2000-08-17 Roland Kreutzer Verfahren und Medikament zur Hemmung der Expression eines vorgegebenen Gens
WO2000063364A2 (en) 1999-04-21 2000-10-26 American Home Products Corporation Methods and compositions for inhibiting the function of polynucleotide sequences
US20040002153A1 (en) * 1999-07-21 2004-01-01 Monia Brett P. Modulation of PTEN expression via oligomeric compounds
GB9925459D0 (en) * 1999-10-27 1999-12-29 Plant Bioscience Ltd Gene silencing
GB9927444D0 (en) 1999-11-19 2000-01-19 Cancer Res Campaign Tech Inhibiting gene expression
US7829693B2 (en) * 1999-11-24 2010-11-09 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of a target gene
DE10160151A1 (de) 2001-01-09 2003-06-26 Ribopharma Ag Verfahren zur Hemmung der Expression eines vorgegebenen Zielgens
DE10100586C1 (de) 2001-01-09 2002-04-11 Ribopharma Ag Verfahren zur Hemmung der Expression eines Ziegens
RU2164944C1 (ru) * 1999-12-09 2001-04-10 Институт молекулярной биологии им. В.А. Энгельгардта РАН Способ изменения генетических свойств организма
US8202979B2 (en) * 2002-02-20 2012-06-19 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid
AU2001260114A1 (en) 2000-03-14 2001-09-24 Syngenta Participations Ag Protoporphyrinogen oxidase ("protox") genes
US20030084471A1 (en) * 2000-03-16 2003-05-01 David Beach Methods and compositions for RNA interference
AU2001245793A1 (en) * 2000-03-16 2001-09-24 Cold Spring Harbor Laboratory Methods and compositions for rna interference
AU2001249622B2 (en) 2000-03-30 2007-06-07 Massachusetts Institute Of Technology RNA sequence-specific mediators of RNA interference
EP2796553B1 (en) 2000-03-30 2019-06-19 Whitehead Institute for Biomedical Research RNA sequence-specific mediators of RNA interference
WO2001092513A1 (en) 2000-05-30 2001-12-06 Johnson & Johnson Research Pty Limited METHODS FOR MEDIATING GENE SUPPRESION BY USING FACTORS THAT ENHANCE RNAi
WO2003103600A2 (en) 2002-06-05 2003-12-18 Invitrogen Corporation Methods and compositions for synthesis of nucleic acid molecules using multiple recognition sites
ES2728168T3 (es) 2000-12-01 2019-10-22 Max Planck Gesellschaft Moléculas pequeñas de ARN que median en la interferencia de ARN
AU2002248173B2 (en) 2000-12-08 2007-04-26 Invitrogen Corporation Compositions and methods for rapidly generating recombinant nucleic acid molecules
JP2004532616A (ja) * 2000-12-28 2004-10-28 ジョンソン・アンド・ジョンソン・リサーチ・ピー・ティー・ワイ・リミテッド 二本鎖rna仲介遺伝子抑制
WO2003035869A1 (de) 2001-10-26 2003-05-01 Ribopharma Ag Verwendung einer doppelsträngigen ribonukleinsäure zur gezielten hemmung der expression eines vorgegebenen zielgens
US7423142B2 (en) * 2001-01-09 2008-09-09 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of anti-apoptotic genes
CA2369944A1 (en) * 2001-01-31 2002-07-31 Nucleonics Inc. Use of post-transcriptional gene silencing for identifying nucleic acid sequences that modulate the function of a cell
CA2442092A1 (en) * 2001-03-26 2002-10-17 Ribozyme Pharmaceuticals, Inc. Oligonucleotide mediated inhibition of hepatitis b virus and hepatitis c virus replication
US20040019001A1 (en) * 2002-02-20 2004-01-29 Mcswiggen James A. RNA interference mediated inhibition of protein typrosine phosphatase-1B (PTP-1B) gene expression using short interfering RNA
US20040006035A1 (en) * 2001-05-29 2004-01-08 Dennis Macejak Nucleic acid mediated disruption of HIV fusogenic peptide interactions
EP1390472A4 (en) * 2001-05-29 2004-11-17 Sirna Therapeutics Inc NUCLEIC ACID TREATMENT OF DISEASES OR SIDES RELATED TO RAS, HER2 AND HIV LEVELS
EP1263250B1 (de) 2001-06-01 2004-03-24 Mobilkom Austria Aktiengesellschaft & Co KG Verfahren zur Bestimmung des Standortes einer Mobilstation in einem Mobilfunksystem
US20030140362A1 (en) * 2001-06-08 2003-07-24 Dennis Macejak In vivo models for screening inhibitors of hepatitis B virus
EP2428571B1 (en) * 2001-09-28 2018-07-18 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. MicroRNA molecules
DE10163098B4 (de) 2001-10-12 2005-06-02 Alnylam Europe Ag Verfahren zur Hemmung der Replikation von Viren
DE10230997A1 (de) * 2001-10-26 2003-07-17 Ribopharma Ag Medikament zur Erhöhung der Wirksamkeit eines Rezeptor-vermittelt Apoptose in Tumorzellen auslösenden Arzneimittels
US20040248835A1 (en) * 2001-10-26 2004-12-09 Anja Krebs Use of a double-stranded ribonucleic acid for treating an infection with a positivestrand rna-virus
US20040121348A1 (en) * 2001-10-26 2004-06-24 Ribopharma Ag Compositions and methods for treating pancreatic cancer
DE10154113A1 (de) 2001-11-03 2003-05-15 Opel Adam Ag Frontstruktur eines Kraftfahrzeuges
DE10202419A1 (de) * 2002-01-22 2003-08-07 Ribopharma Ag Verfahren zur Hemmung der Expression eines durch eine Chromosomen-Aberration entstandenen Zielgens
WO2003064621A2 (en) 2002-02-01 2003-08-07 Ambion, Inc. HIGH POTENCY siRNAS FOR REDUCING THE EXPRESSION OF TARGET GENES
JP2005527198A (ja) * 2002-02-14 2005-09-15 シティ・オブ・ホープ 哺乳動物細胞において、干渉rna分子を産生する方法、および該干渉rna分子の療法的使用
US20040005593A1 (en) * 2002-03-06 2004-01-08 Rigel Pharmaceuticals, Inc. Novel method for delivery and intracellular synthesis of siRNA molecules
EP1495141A4 (en) * 2002-03-20 2006-03-22 Massachusetts Inst Technology HIV THERAPEUTIC
US20030180756A1 (en) * 2002-03-21 2003-09-25 Yang Shi Compositions and methods for suppressing eukaryotic gene expression
US20040053876A1 (en) * 2002-03-26 2004-03-18 The Regents Of The University Of Michigan siRNAs and uses therof
AU2003237686A1 (en) 2002-05-24 2003-12-12 Max-Planck Gesellschaft Zur Forderung Der Wissenschaften E.V. Rna interference mediating small rna molecules
GB2406169B (en) 2002-06-12 2006-11-01 Ambion Inc Methods and compositions relating to labeled rna molecules that reduce gene expression
EP2333062A1 (en) 2002-07-10 2011-06-15 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. RNA-interference by single-stranded RNA molecules
WO2004015107A2 (en) 2002-08-05 2004-02-19 Atugen Ag Further novel forms of interfering rna molecules
US20040241854A1 (en) * 2002-08-05 2004-12-02 Davidson Beverly L. siRNA-mediated gene silencing
WO2004014933A1 (en) 2002-08-07 2004-02-19 University Of Massachusetts Compositions for rna interference and methods of use thereof
WO2004027030A2 (en) 2002-09-18 2004-04-01 Isis Pharmaceuticals, Inc. Efficient reduction of target rna’s by single- and double-stranded oligomeric compounds
EP1556402B1 (en) 2002-09-25 2011-06-22 University of Massachusetts In vivo gene silencing by chemically modified and stable sirna
WO2004044139A2 (en) 2002-11-05 2004-05-27 Isis Parmaceuticals, Inc. Modified oligonucleotides for use in rna interference
PT2284266E (pt) 2002-11-14 2013-12-17 Thermo Fisher Scient Biosciences Inc Siarn contra tp53
AU2003295539A1 (en) 2002-11-15 2004-06-15 University Of Massachusetts Allele-targeted rna interference
WO2004047764A2 (en) * 2002-11-22 2004-06-10 University Of Massachusetts Modulation of hiv replication by rna interference
WO2004063375A1 (en) 2003-01-15 2004-07-29 Hans Prydz OPTIMIZING siRNA BY RNAi ANTISENSE
US20040224328A1 (en) * 2003-01-15 2004-11-11 Hans Prydz siRNA screening method
WO2004065600A2 (en) 2003-01-17 2004-08-05 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Rna interference by palindromic or modified rna molecules
EP1583832B1 (en) 2003-01-17 2010-12-01 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Inducible small interfering rna (sirna) expression constructs for targeted gene silencing
KR20050115231A (ko) 2003-02-10 2005-12-07 내셔날 인스티튜트 오브 어드밴스드 인더스트리얼 사이언스 앤드 테크놀로지 포유동물 세포의 조절
ATE528402T1 (de) * 2003-02-19 2011-10-15 Commw Scient Ind Res Org Effiziente genabschaltung in pflanzen unter verwendung von kurzen dsrna-sequenzen
PT1633767T (pt) 2003-06-02 2019-02-27 Univ Massachusetts Métodos e composições para controlar a eficácia do silenciamento de arn
US6998203B2 (en) * 2003-08-01 2006-02-14 Intel Corporation Proximity correcting lithography mask blanks
US7507809B2 (en) 2005-01-07 2009-03-24 Alnylam Pharmaceuticals, Inc. RNAi modulation of RSV and therapeutic uses thereof
JP6348063B2 (ja) 2011-07-08 2018-06-27 インテヴァ プロダクツ, エルエルシーInteva Products, Llc 車両内装部材にステッチを施す方法

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Publication number Priority date Publication date Assignee Title
CN101213300B (zh) * 2005-06-01 2013-01-23 聚加转染股份有限公司 用于rna干扰的寡核苷酸及其生物学应用
CN100445381C (zh) * 2005-06-10 2008-12-24 中国人民解放军军事医学科学院基础医学研究所 带有单链polyA尾巴的siRNA分子制备方法和应用
US10633654B2 (en) 2008-02-11 2020-04-28 Phio Pharmaceuticals Corp. Modified RNAi polynucleotides and uses thereof
US10131904B2 (en) 2008-02-11 2018-11-20 Rxi Pharmaceuticals Corporation Modified RNAi polynucleotides and uses thereof
US8815818B2 (en) 2008-07-18 2014-08-26 Rxi Pharmaceuticals Corporation Phagocytic cell delivery of RNAI
US10138485B2 (en) 2008-09-22 2018-11-27 Rxi Pharmaceuticals Corporation Neutral nanotransporters
US10774330B2 (en) 2008-09-22 2020-09-15 Phio Pharmaceuticals Corp. Reduced size self-delivering RNAI compounds
US8796443B2 (en) 2008-09-22 2014-08-05 Rxi Pharmaceuticals Corporation Reduced size self-delivering RNAi compounds
US11396654B2 (en) 2008-09-22 2022-07-26 Phio Pharmaceuticals Corp. Neutral nanotransporters
US10041073B2 (en) 2008-09-22 2018-08-07 Rxi Pharmaceuticals Corporation Reduced size self-delivering RNAi compounds
US9175289B2 (en) 2008-09-22 2015-11-03 Rxi Pharmaceuticals Corporation Reduced size self-delivering RNAi compounds
US9303259B2 (en) 2008-09-22 2016-04-05 Rxi Pharmaceuticals Corporation RNA interference in skin indications
US10876119B2 (en) 2008-09-22 2020-12-29 Phio Pharmaceuticals Corp. Reduced size self-delivering RNAI compounds
US10815485B2 (en) 2008-09-22 2020-10-27 Phio Pharmaceuticals Corp. RNA interference in skin indications
US9938530B2 (en) 2008-09-22 2018-04-10 Rxi Pharmaceuticals Corporation RNA interference in skin indications
US8664189B2 (en) 2008-09-22 2014-03-04 Rxi Pharmaceuticals Corporation RNA interference in skin indications
US11254940B2 (en) 2008-11-19 2022-02-22 Phio Pharmaceuticals Corp. Inhibition of MAP4K4 through RNAi
US9074211B2 (en) 2008-11-19 2015-07-07 Rxi Pharmaceuticals Corporation Inhibition of MAP4K4 through RNAI
US9493774B2 (en) 2009-01-05 2016-11-15 Rxi Pharmaceuticals Corporation Inhibition of PCSK9 through RNAi
US10167471B2 (en) 2009-01-05 2019-01-01 Rxi Pharmaceuticals Corporation Inhibition of PCSK9 through RNAI
US10479992B2 (en) 2009-02-04 2019-11-19 Phio Pharmaceuticals Corp. RNA duplexes with single stranded phosphorothioate nucleotide regions for additional functionality
US11667915B2 (en) 2009-02-04 2023-06-06 Phio Pharmaceuticals Corp. RNA duplexes with single stranded phosphorothioate nucleotide regions for additional functionality
US9745574B2 (en) 2009-02-04 2017-08-29 Rxi Pharmaceuticals Corporation RNA duplexes with single stranded phosphorothioate nucleotide regions for additional functionality
CN102333869A (zh) * 2009-02-24 2012-01-25 里博克斯艾克斯有限公司 小干扰rna的改进设计
US11118178B2 (en) 2010-03-24 2021-09-14 Phio Pharmaceuticals Corp. Reduced size self-delivering RNAI compounds
US9963702B2 (en) 2010-03-24 2018-05-08 Rxi Pharmaceuticals Corporation RNA interference in dermal and fibrotic indications
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US10240149B2 (en) 2010-03-24 2019-03-26 Phio Pharmaceuticals Corp. Reduced size self-delivering RNAi compounds
US10184124B2 (en) 2010-03-24 2019-01-22 Phio Pharmaceuticals Corp. RNA interference in ocular indications
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