WO2011091324A8 - Methods of generating zinc finger nucleases having altered activity - Google Patents

Methods of generating zinc finger nucleases having altered activity Download PDF

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Publication number
WO2011091324A8
WO2011091324A8 PCT/US2011/022154 US2011022154W WO2011091324A8 WO 2011091324 A8 WO2011091324 A8 WO 2011091324A8 US 2011022154 W US2011022154 W US 2011022154W WO 2011091324 A8 WO2011091324 A8 WO 2011091324A8
Authority
WO
WIPO (PCT)
Prior art keywords
methods
zinc finger
catalytic activity
zfn
altered
Prior art date
Application number
PCT/US2011/022154
Other languages
French (fr)
Other versions
WO2011091324A2 (en
WO2011091324A3 (en
Inventor
III Carlos F. BARBAS
Jing Guo
Original Assignee
The Scripps Research Institute
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 The Scripps Research Institute filed Critical The Scripps Research Institute
Priority to EP11735277.3A priority Critical patent/EP2526199A4/en
Priority to US13/574,223 priority patent/US20120329067A1/en
Publication of WO2011091324A2 publication Critical patent/WO2011091324A2/en
Publication of WO2011091324A3 publication Critical patent/WO2011091324A3/en
Publication of WO2011091324A8 publication Critical patent/WO2011091324A8/en

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    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • C07K14/4703—Inhibitors; Suppressors
    • 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
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases RNAses, DNAses
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
    • G01N2333/922—Ribonucleases (RNAses); Deoxyribonucleases (DNAses)

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Plant Pathology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Provided herein are zinc linger nucleases having altered, arid in particular, improved catalytic activity and methods of generating such nucleases. Accordingly, there are provided methods for identifying improved catalytic activity of a ZFN by expressing a mutated zinc finger nuclease in a cell containing a reporter construct with a toxic gene, and a zinc finger nuclease cleavage site that is recognized by the ZFN. Survival of the cell is positively correlated with catalytic activity of the ZFN; thus, libraries of mutated ZFKs may be selected for altered catalytic activity based on relative survival rates, Methods of using identified ZFNs are also provided.
PCT/US2011/022154 2010-01-22 2011-01-21 Methods of generating zinc finger nucleases having altered activity WO2011091324A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP11735277.3A EP2526199A4 (en) 2010-01-22 2011-01-21 Methods of generating zinc finger nucleases having altered activity
US13/574,223 US20120329067A1 (en) 2010-01-22 2011-01-21 Methods of Generating Zinc Finger Nucleases Having Altered Activity

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US29759810P 2010-01-22 2010-01-22
US61/297,598 2010-01-22
US32903510P 2010-04-28 2010-04-28
US61/329,035 2010-04-28

Publications (3)

Publication Number Publication Date
WO2011091324A2 WO2011091324A2 (en) 2011-07-28
WO2011091324A3 WO2011091324A3 (en) 2011-12-22
WO2011091324A8 true WO2011091324A8 (en) 2012-08-30

Family

ID=44307625

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2011/022154 WO2011091324A2 (en) 2010-01-22 2011-01-21 Methods of generating zinc finger nucleases having altered activity

Country Status (2)

Country Link
EP (1) EP2526199A4 (en)
WO (1) WO2011091324A2 (en)

Families Citing this family (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102660642A (en) * 2012-04-24 2012-09-12 西北农林科技大学 Screening system for screening zinc finger protein
WO2016176617A2 (en) 2015-04-29 2016-11-03 New York University Method for treating high-grade gliomas
US20210047633A1 (en) * 2017-03-29 2021-02-18 Editas Medicine, Inc. Selection methods
WO2018227114A1 (en) 2017-06-09 2018-12-13 Editas Medicine, Inc. Engineered cas9 nucleases
CN108118059B (en) * 2017-12-30 2021-03-19 苏州金唯智生物科技有限公司 Improved promoter, vector composed of improved promoter and application of improved promoter
JP2022547699A (en) * 2019-09-12 2022-11-15 グラクソスミスクライン、インテレクチュアル、プロパティー、ディベロップメント、リミテッド Library screening method
EP4143304A2 (en) 2020-04-28 2023-03-08 Intellia Therapeutics, Inc. Methods of in vitro cell delivery
EP4242237A1 (en) * 2020-11-06 2023-09-13 Editforce, Inc. Foki nuclease domain variant
TW202239959A (en) 2020-12-23 2022-10-16 美商英特利亞醫療公司 Compositions and methods for reducing hla-a in a cell
JP2024501246A (en) 2020-12-23 2024-01-11 インテリア セラピューティクス，インコーポレイテッド Compositions and methods for genetically modifying CIITA in cells
CA3206484A1 (en) 2020-12-30 2022-07-07 Intellia Therapeutics, Inc. Engineered t cells
CR20230534A (en) 2021-04-17 2024-02-12 Intellia Therapeutics Inc Lipid nanoparticle compositions
IL307740A (en) 2021-04-17 2023-12-01 Intellia Therapeutics Inc Inhibitors of dna-dependent protein kinase and compositions and uses thereof
AU2022257050A1 (en) 2021-04-17 2023-11-30 Intellia Therapeutics, Inc. Lipid nanoparticle compositions
WO2023205148A1 (en) 2022-04-19 2023-10-26 Intellia Therapeutics, Inc. Chimeric antigen receptor compositions and uses
WO2023245108A2 (en) 2022-06-16 2023-12-21 Intellia Therapeutics, Inc. Compositions and methods for reducing mhc class i in a cell
WO2024006955A1 (en) 2022-06-29 2024-01-04 Intellia Therapeutics, Inc. Engineered t cells

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003087341A2 (en) * 2002-01-23 2003-10-23 The University Of Utah Research Foundation Targeted chromosomal mutagenesis using zinc finger nucleases
US20090042186A1 (en) * 2005-05-06 2009-02-12 Alexander Mankin Mapping new sites for antibiotic action in the ribosome
DE602007005634D1 (en) * 2006-05-25 2010-05-12 Sangamo Biosciences Inc VARIANT FOKI CREVICE HOLLAND DOMAINS
EP2568048A1 (en) * 2007-06-29 2013-03-13 Pioneer Hi-Bred International, Inc. Methods for altering the genome of a monocot plant cell

Also Published As

Publication number Publication date
EP2526199A2 (en) 2012-11-28
WO2011091324A2 (en) 2011-07-28
WO2011091324A3 (en) 2011-12-22
EP2526199A4 (en) 2013-08-07

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