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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
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/US2011/022154
Other languages
French (fr)
Other versions
WO2011091324A2 (en
WO2011091324A3 (en
Inventor
III Carlos F. BARBAS
Jing Guo
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Scripps Research Institute
Original Assignee
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 Scripps Research Institute filed Critical 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
Anticipated expiration legal-status Critical
Publication of WO2011091324A8 publication Critical patent/WO2011091324A8/en
Ceased legal-status Critical Current

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
    • 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 [RNase]; Deoxyribonucleases [DNase]
    • 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)

Landscapes

  • 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)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Computational Biology (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 Ceased WO2011091324A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP11735277.3A EP2526199A4 (en) 2010-01-22 2011-01-21 METHODS FOR PRODUCING ZINC FINGER NUCLEASES HAVING MODIFIED 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 Ceased 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 (25)

* 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
EP3289104B1 (en) 2015-04-29 2020-11-04 New York University Method for treating high-grade gliomas
WO2018183766A1 (en) * 2017-03-29 2018-10-04 Editas Medicine, Inc. Selection methods
EP3635104A1 (en) 2017-06-09 2020-04-15 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 An improved promoter and its composition vector and application
EP4028585A1 (en) * 2019-09-12 2022-07-20 GlaxoSmithKline Intellectual Property Development Limited Method for screening libraries
CN116018403A (en) 2020-04-28 2023-04-25 因特利亚治疗公司 In vitro cell delivery methods
JP7752342B2 (en) * 2020-11-06 2025-10-10 エディットフォース株式会社 Mutants of the FokI nuclease domain
CA3206284A1 (en) 2020-12-23 2022-06-30 Intellia Therapeutics, Inc. Compositions and methods for reducing hla-a in a cell
WO2022140587A1 (en) 2020-12-23 2022-06-30 Intellia Therapeutics, Inc. Compositions and methods for genetically modifying ciita in a cell
AU2021411521A1 (en) 2020-12-30 2023-08-03 Intellia Therapeutics, Inc. Engineered t cells
WO2022221696A1 (en) 2021-04-17 2022-10-20 Intellia Therapeutics, Inc. Inhibitors of dna-dependent protein kinase and compositions and uses thereof
JP2024515647A (en) 2021-04-17 2024-04-10 インテリア セラピューティクス，インコーポレーテッド Lipid Nanoparticle Compositions
US20250090471A1 (en) 2021-04-17 2025-03-20 Intellia Therapeutics, Inc. Lipid nanoparticle compositions
US20250268940A1 (en) 2022-04-19 2025-08-28 Intellia Therapeutics, Inc. Chimeric antigen receptor compositions and uses
AU2023293131A1 (en) 2022-06-16 2024-12-12 Intellia Therapeutics, Inc. Compositions and methods for reducing mhc class i in a cell
IL317790A (en) 2022-06-29 2025-02-01 Intellia Therapeutics Inc Engineered t cells
AU2024270788A1 (en) 2023-05-18 2024-11-21 Clade Therapeutics, Inc. Inhibiting natural killer cell cytotoxicity against cell therapies
TW202502729A (en) 2023-05-19 2025-01-16 美商英特利亞醫療公司 Ionizable amine lipids
TW202521564A (en) 2023-08-14 2025-06-01 美商英特利亞醫療公司 Cd70 car-t compositions and methods for cell-based therapy
WO2025038637A1 (en) 2023-08-14 2025-02-20 Intellia Therapeutics, Inc. Compositions and methods for genetically modifying transforming growth factor beta receptor type 2 (tgfβr2)
TW202515994A (en) 2023-08-14 2025-04-16 美商英特利亞醫療公司 Compositions and methods for genetically modifying cd70
WO2025038648A1 (en) 2023-08-14 2025-02-20 Intellia Therapeutics, Inc. Compositions and methods for genetically modifying transforming growth factor beta receptor type 2 (tgfβr2)
WO2025049481A1 (en) 2023-08-28 2025-03-06 Intellia Therapeutics, Inc. Methods of editing an hla-a gene in vitro
WO2025240946A1 (en) 2024-05-17 2025-11-20 Intellia Therapeutics, Inc. Lipid nanoparticles and lipid nanoparticle compositions

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8106255B2 (en) * 2002-01-23 2012-01-31 Dana Carroll Targeted chromosomal mutagenasis using zinc finger nucleases
US20080131962A1 (en) * 2006-05-25 2008-06-05 Sangamo Biosciences, Inc. Engineered cleavage half-domains
US20090042186A1 (en) * 2005-05-06 2009-02-12 Alexander Mankin Mapping new sites for antibiotic action in the ribosome
US8912392B2 (en) * 2007-06-29 2014-12-16 Pioneer Hi-Bred International, Inc. Methods for altering the genome of a monocot plant cell

Also Published As

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

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