WO2008005529A3 - Cell-mediated directed evolution - Google Patents

Cell-mediated directed evolution Download PDF

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Publication number
WO2008005529A3
WO2008005529A3 PCT/US2007/015568 US2007015568W WO2008005529A3 WO 2008005529 A3 WO2008005529 A3 WO 2008005529A3 US 2007015568 W US2007015568 W US 2007015568W WO 2008005529 A3 WO2008005529 A3 WO 2008005529A3
Authority
WO
WIPO (PCT)
Prior art keywords
mutants
cell
interest
directed evolution
select
Prior art date
Application number
PCT/US2007/015568
Other languages
French (fr)
Other versions
WO2008005529A2 (en
Inventor
Virginia W Cornish
Pamela Peralta-Yahya
Jonathan Bronson
Original Assignee
Trustees Columbia University I
Virginia W Cornish
Pamela Peralta-Yahya
Jonathan Bronson
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Trustees Columbia University I, Virginia W Cornish, Pamela Peralta-Yahya, Jonathan Bronson filed Critical Trustees Columbia University I
Publication of WO2008005529A2 publication Critical patent/WO2008005529A2/en
Publication of WO2008005529A3 publication Critical patent/WO2008005529A3/en
Priority to US12/348,693 priority Critical patent/US9169481B2/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1058—Directional evolution of libraries, e.g. evolution of libraries is achieved by mutagenesis and screening or selection of mixed population of organisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1055—Protein x Protein interaction, e.g. two hybrid selection
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1086—Preparation or screening of expression libraries, e.g. reporter assays

Abstract

The present invention relates to systems which harness the molecular biology of a living cell to direct evolution of biological entities of interest. According to the invention, a host cell is engineered to facilitate mutation of a nucleic acid target corresponding to that entity and select for desirable mutants. As applied to populations of host cells, the invention provides a means to generate and contemporaneously select mutants of interest, allowing for the production of extremely diverse libraries enriched in the most 'fit' mutants.
PCT/US2007/015568 2006-07-07 2007-07-05 Cell-mediated directed evolution WO2008005529A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US12/348,693 US9169481B2 (en) 2006-07-07 2009-01-05 Cell-mediated directed evolution

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US81953106P 2006-07-07 2006-07-07
US81953206P 2006-07-07 2006-07-07
US60/819,532 2006-07-07
US60/819,531 2006-07-07
US89183307P 2007-02-27 2007-02-27
US60/891,833 2007-02-27

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US12/348,693 Continuation US9169481B2 (en) 2006-07-07 2009-01-05 Cell-mediated directed evolution

Publications (2)

Publication Number Publication Date
WO2008005529A2 WO2008005529A2 (en) 2008-01-10
WO2008005529A3 true WO2008005529A3 (en) 2008-09-18

Family

ID=38895234

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2007/015568 WO2008005529A2 (en) 2006-07-07 2007-07-05 Cell-mediated directed evolution

Country Status (2)

Country Link
US (1) US9169481B2 (en)
WO (1) WO2008005529A2 (en)

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CA3073384A1 (en) 2008-09-05 2010-03-11 President And Fellows Of Harvard College Continuous directed evolution of proteins and nucleic acids
EP3202903B1 (en) 2010-12-22 2020-02-12 President and Fellows of Harvard College Continuous directed evolution
US20150044192A1 (en) 2013-08-09 2015-02-12 President And Fellows Of Harvard College Methods for identifying a target site of a cas9 nuclease
US9340800B2 (en) 2013-09-06 2016-05-17 President And Fellows Of Harvard College Extended DNA-sensing GRNAS
US10179911B2 (en) 2014-01-20 2019-01-15 President And Fellows Of Harvard College Negative selection and stringency modulation in continuous evolution systems
CA2956224A1 (en) 2014-07-30 2016-02-11 President And Fellows Of Harvard College Cas9 proteins including ligand-dependent inteins
WO2016077052A2 (en) 2014-10-22 2016-05-19 President And Fellows Of Harvard College Evolution of proteases
US11299729B2 (en) 2015-04-17 2022-04-12 President And Fellows Of Harvard College Vector-based mutagenesis system
JP6586659B2 (en) 2015-05-19 2019-10-09 本田技研工業株式会社 Thermostable glycoside hydrolase
US10392674B2 (en) 2015-07-22 2019-08-27 President And Fellows Of Harvard College Evolution of site-specific recombinases
US11524983B2 (en) 2015-07-23 2022-12-13 President And Fellows Of Harvard College Evolution of Bt toxins
WO2017019895A1 (en) 2015-07-30 2017-02-02 President And Fellows Of Harvard College Evolution of talens
IL310721A (en) 2015-10-23 2024-04-01 President And Fellows Of Harvard College Nucleobase editors and uses thereof
EP3192866A1 (en) * 2016-01-15 2017-07-19 CIC nanoGUNE - Asociación Centro de Investigación Cooperativa en Nanociencias Endocellulases and uses thereof
KR20230095129A (en) 2016-08-03 2023-06-28 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 Adenosine nucleobase editors and uses thereof
CN109804066A (en) 2016-08-09 2019-05-24 哈佛大学的校长及成员们 Programmable CAS9- recombination enzyme fusion proteins and application thereof
US11542509B2 (en) 2016-08-24 2023-01-03 President And Fellows Of Harvard College Incorporation of unnatural amino acids into proteins using base editing
KR20240007715A (en) 2016-10-14 2024-01-16 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 Aav delivery of nucleobase editors
WO2018119359A1 (en) 2016-12-23 2018-06-28 President And Fellows Of Harvard College Editing of ccr5 receptor gene to protect against hiv infection
US11898179B2 (en) 2017-03-09 2024-02-13 President And Fellows Of Harvard College Suppression of pain by gene editing
WO2018165629A1 (en) 2017-03-10 2018-09-13 President And Fellows Of Harvard College Cytosine to guanine base editor
IL269458B2 (en) 2017-03-23 2024-02-01 Harvard College Nucleobase editors comprising nucleic acid programmable dna binding proteins
US11560566B2 (en) 2017-05-12 2023-01-24 President And Fellows Of Harvard College Aptazyme-embedded guide RNAs for use with CRISPR-Cas9 in genome editing and transcriptional activation
US11447809B2 (en) 2017-07-06 2022-09-20 President And Fellows Of Harvard College Evolution of tRNA synthetases
US11732274B2 (en) 2017-07-28 2023-08-22 President And Fellows Of Harvard College Methods and compositions for evolving base editors using phage-assisted continuous evolution (PACE)
EP3676376A2 (en) 2017-08-30 2020-07-08 President and Fellows of Harvard College High efficiency base editors comprising gam
US11624130B2 (en) 2017-09-18 2023-04-11 President And Fellows Of Harvard College Continuous evolution for stabilized proteins
US11795443B2 (en) 2017-10-16 2023-10-24 The Broad Institute, Inc. Uses of adenosine base editors
WO2019241649A1 (en) 2018-06-14 2019-12-19 President And Fellows Of Harvard College Evolution of cytidine deaminases
WO2020191248A1 (en) 2019-03-19 2020-09-24 The Broad Institute, Inc. Method and compositions for editing nucleotide sequences
EP3872173A1 (en) 2020-02-28 2021-09-01 CIC nanoGUNE - Asociación Centro de Investigación Cooperativa en Nanociencias Method for producing crystalline nanocellulose
JP2023525304A (en) 2020-05-08 2023-06-15 ザ ブロード インスティテュート，インコーポレーテッド Methods and compositions for simultaneous editing of both strands of a target double-stranded nucleotide sequence

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Also Published As

Publication number Publication date
WO2008005529A2 (en) 2008-01-10
US20090209038A1 (en) 2009-08-20
US9169481B2 (en) 2015-10-27

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