EA037825B1 - ВЫСОКОТОЧНЫЕ ВАРИАНТЫ Cas9 И ИХ ПРИМЕНЕНИЯ - Google Patents

ВЫСОКОТОЧНЫЕ ВАРИАНТЫ Cas9 И ИХ ПРИМЕНЕНИЯ Download PDF

Info

Publication number
EA037825B1
EA037825B1 EA201991913A EA201991913A EA037825B1 EA 037825 B1 EA037825 B1 EA 037825B1 EA 201991913 A EA201991913 A EA 201991913A EA 201991913 A EA201991913 A EA 201991913A EA 037825 B1 EA037825 B1 EA 037825B1
Authority
EA
Eurasian Patent Office
Prior art keywords
mutations
cas9
seq
target
spcas9
Prior art date
Application number
EA201991913A
Other languages
English (en)
Russian (ru)
Other versions
EA201991913A1 (ru
Inventor
Анна Черезето
Антонио Казини
Джанлука Петрис
Альберто Инга
Микеле Оливьери
Original Assignee
Университа` Дельи Студи Ди Тренто
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 Университа` Дельи Студи Ди Тренто filed Critical Университа` Дельи Студи Ди Тренто
Publication of EA201991913A1 publication Critical patent/EA201991913A1/ru
Publication of EA037825B1 publication Critical patent/EA037825B1/ru

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
    • 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]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/71—Fusion polypeptide containing domain for protein-protein interaction containing domain for transcriptional activaation, e.g. VP16
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/80—Fusion polypeptide containing a DNA binding domain, e.g. Lacl or Tet-repressor
    • 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
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/80—Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
EA201991913A 2017-02-14 2018-02-14 ВЫСОКОТОЧНЫЕ ВАРИАНТЫ Cas9 И ИХ ПРИМЕНЕНИЯ EA037825B1 (ru)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IT102017000016321A IT201700016321A1 (it) 2017-02-14 2017-02-14 Mutanti di cas9 ad alta specificita' e loro applicazioni.
PCT/EP2018/053717 WO2018149888A1 (en) 2017-02-14 2018-02-14 High-fidelity cas9 variants and applications thereof

Publications (2)

Publication Number Publication Date
EA201991913A1 EA201991913A1 (ru) 2020-01-22
EA037825B1 true EA037825B1 (ru) 2021-05-25

Family

ID=59067822

Family Applications (1)

Application Number Title Priority Date Filing Date
EA201991913A EA037825B1 (ru) 2017-02-14 2018-02-14 ВЫСОКОТОЧНЫЕ ВАРИАНТЫ Cas9 И ИХ ПРИМЕНЕНИЯ

Country Status (12)

Country Link
US (1) US11525127B2 (https=)
EP (1) EP3583209B8 (https=)
JP (1) JP7232531B2 (https=)
KR (1) KR20190116407A (https=)
CN (1) CN110520528B (https=)
AU (1) AU2018221644A1 (https=)
BR (1) BR112019016765A2 (https=)
CA (1) CA3053218A1 (https=)
EA (1) EA037825B1 (https=)
IT (1) IT201700016321A1 (https=)
MX (1) MX2019009651A (https=)
WO (1) WO2018149888A1 (https=)

Families Citing this family (66)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP6261500B2 (ja) 2011-07-22 2018-01-17 プレジデント アンド フェローズ オブ ハーバード カレッジ ヌクレアーゼ切断特異性の評価および改善
US20150044192A1 (en) 2013-08-09 2015-02-12 President And Fellows Of Harvard College Methods for identifying a target site of a cas9 nuclease
US9359599B2 (en) 2013-08-22 2016-06-07 President And Fellows Of Harvard College Engineered transcription activator-like effector (TALE) domains and uses thereof
US9388430B2 (en) 2013-09-06 2016-07-12 President And Fellows Of Harvard College Cas9-recombinase fusion proteins and uses thereof
US9526784B2 (en) 2013-09-06 2016-12-27 President And Fellows Of Harvard College Delivery system for functional nucleases
US9340799B2 (en) 2013-09-06 2016-05-17 President And Fellows Of Harvard College MRNA-sensing switchable gRNAs
US20150165054A1 (en) 2013-12-12 2015-06-18 President And Fellows Of Harvard College Methods for correcting caspase-9 point mutations
WO2016022363A2 (en) 2014-07-30 2016-02-11 President And Fellows Of Harvard College Cas9 proteins including ligand-dependent inteins
EP3204496A1 (en) 2014-10-10 2017-08-16 Editas Medicine, Inc. Compositions and methods for promoting homology directed repair
CA2963820A1 (en) 2014-11-07 2016-05-12 Editas Medicine, Inc. Methods for improving crispr/cas-mediated genome-editing
AU2016261358B2 (en) 2015-05-11 2021-09-16 Editas Medicine, Inc. Optimized CRISPR/Cas9 systems and methods for gene editing in stem cells
EP3307887A1 (en) 2015-06-09 2018-04-18 Editas Medicine, Inc. Crispr/cas-related methods and compositions for improving transplantation
AU2016326711B2 (en) 2015-09-24 2022-11-03 Editas Medicine, Inc. Use of exonucleases to improve CRISPR/Cas-mediated genome editing
JP7109784B2 (ja) 2015-10-23 2022-08-01 プレジデント アンド フェローズ オブ ハーバード カレッジ 遺伝子編集のための進化したＣａｓ９蛋白質
EP3433363A1 (en) 2016-03-25 2019-01-30 Editas Medicine, Inc. Genome editing systems comprising repair-modulating enzyme molecules and methods of their use
WO2017180694A1 (en) 2016-04-13 2017-10-19 Editas Medicine, Inc. Cas9 fusion molecules gene editing systems, and methods of use thereof
KR102547316B1 (ko) 2016-08-03 2023-06-23 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 아데노신 핵염기 편집제 및 그의 용도
US11661590B2 (en) 2016-08-09 2023-05-30 President And Fellows Of Harvard College Programmable CAS9-recombinase fusion proteins and uses 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
CN118726313A (zh) 2016-10-07 2024-10-01 综合Dna技术公司 化脓链球菌cas9突变基因和由其编码的多肽
US11242542B2 (en) * 2016-10-07 2022-02-08 Integrated Dna Technologies, Inc. S. pyogenes Cas9 mutant genes and polypeptides encoded by same
EP3526320A1 (en) 2016-10-14 2019-08-21 President and Fellows of Harvard College Aav delivery of nucleobase editors
US11377645B2 (en) 2016-12-16 2022-07-05 The Rockefeller University Compositions and methods using Cas9 with enhanced spacer acquisition function
WO2018119010A1 (en) 2016-12-19 2018-06-28 Editas Medicine, Inc. Assessing nuclease cleavage
WO2018119359A1 (en) 2016-12-23 2018-06-28 President And Fellows Of Harvard College Editing of ccr5 receptor gene to protect against hiv infection
EP3565907B1 (en) 2017-01-06 2022-05-04 Editas Medicine, Inc. Methods of assessing nuclease cleavage
IT201700016321A1 (it) 2017-02-14 2018-08-14 Univ Degli Studi Di Trento Mutanti di cas9 ad alta specificita' e loro applicazioni.
JP2020510038A (ja) 2017-03-09 2020-04-02 プレジデント アンド フェローズ オブ ハーバード カレッジ がんワクチン
EP3592853A1 (en) 2017-03-09 2020-01-15 President and Fellows of Harvard College Suppression of pain by gene editing
JP2020510439A (ja) 2017-03-10 2020-04-09 プレジデント アンド フェローズ オブ ハーバード カレッジ シトシンからグアニンへの塩基編集因子
CA3057192A1 (en) 2017-03-23 2018-09-27 President And Fellows Of Harvard College Nucleobase editors comprising nucleic acid programmable dna binding proteins
US11499151B2 (en) 2017-04-28 2022-11-15 Editas Medicine, Inc. Methods and systems for analyzing guide RNA molecules
WO2018209320A1 (en) 2017-05-12 2018-11-15 President And Fellows Of Harvard College Aptazyme-embedded guide rnas for use with crispr-cas9 in genome editing and transcriptional activation
WO2018227114A1 (en) 2017-06-09 2018-12-13 Editas Medicine, Inc. Engineered cas9 nucleases
US11866726B2 (en) 2017-07-14 2024-01-09 Editas Medicine, Inc. Systems and methods for targeted integration and genome editing and detection thereof using integrated priming sites
WO2019023680A1 (en) 2017-07-28 2019-01-31 President And Fellows Of Harvard College METHODS AND COMPOSITIONS FOR EVOLUTION OF BASIC EDITORS USING PHAGE-ASSISTED CONTINUOUS EVOLUTION (PACE)
EP3676376B1 (en) 2017-08-30 2025-01-15 President and Fellows of Harvard College High efficiency base editors comprising gam
KR20250107288A (ko) 2017-10-16 2025-07-11 더 브로드 인스티튜트, 인코퍼레이티드 아데노신 염기 편집제의 용도
WO2019118949A1 (en) 2017-12-15 2019-06-20 The Broad Institute, Inc. Systems and methods for predicting repair outcomes in genetic engineering
EP3797160A1 (en) 2018-05-23 2021-03-31 The Broad Institute Inc. Base editors and uses thereof
EP3823633A4 (en) 2018-06-29 2023-05-03 Editas Medicine, Inc. SYNTHETIC LEAD MOLECULES, COMPOSITIONS AND METHODS RELATED THERETO
US12522807B2 (en) 2018-07-09 2026-01-13 The Broad Institute, Inc. RNA programmable epigenetic RNA modifiers and uses thereof
KR20210060541A (ko) * 2018-09-19 2021-05-26 더 유니버시티 오브 홍콩 개선된 고처리량 조합 유전적 변형 시스템 및 최적화된 Cas9 효소 변이체
WO2020092453A1 (en) 2018-10-29 2020-05-07 The Broad Institute, Inc. Nucleobase editors comprising geocas9 and uses thereof
US12351837B2 (en) 2019-01-23 2025-07-08 The Broad Institute, Inc. Supernegatively charged proteins and uses thereof
WO2020160125A1 (en) * 2019-01-29 2020-08-06 Flagship Pioneering Innovations V, Inc. Compositions comprising an endonuclease and methods for purifying an endonuclease
JP7657726B2 (ja) 2019-03-19 2025-04-07 ザ ブロード インスティテュート，インコーポレーテッド 編集ヌクレオチド配列を編集するための方法および組成物
US12473543B2 (en) 2019-04-17 2025-11-18 The Broad Institute, Inc. Adenine base editors with reduced off-target effects
WO2020257325A1 (en) 2019-06-17 2020-12-24 Vertex Pharmaceuticals Inc. Compositions and methods for editing beta-globin for treatment of hemaglobinopathies
US12435330B2 (en) 2019-10-10 2025-10-07 The Broad Institute, Inc. Methods and compositions for prime editing RNA
US20230174972A1 (en) * 2019-10-15 2023-06-08 Agency For Science, Technology And Research Assays For Measuring Nucleic Acid Modifying Enzyme Activity
WO2021122944A1 (en) 2019-12-18 2021-06-24 Alia Therapeutics Srl Compositions and methods for treating retinitis pigmentosa
CA3163463A1 (en) * 2020-03-11 2021-09-16 Fuqiang Chen High fidelity spcas9 nucleases for genome modification
DE112021002672T5 (de) 2020-05-08 2023-04-13 President And Fellows Of Harvard College Vefahren und zusammensetzungen zum gleichzeitigen editieren beider stränge einer doppelsträngigen nukleotid-zielsequenz
WO2022018638A1 (en) 2020-07-21 2022-01-27 Crispr Therapeutics Ag Genome-editing compositions and methods to modulate faah for treatment of neurological disorders
AU2021400745A1 (en) 2020-12-17 2023-07-20 Vertex Pharmaceuticals Incorporated Compositions and methods for editing beta-globin for treatment of hemaglobinopathies
CN112680430B (zh) * 2020-12-28 2023-06-06 南方医科大学 一种CRISPR SpCas9突变体及其应用
CN113005141A (zh) * 2021-01-05 2021-06-22 温州医科大学 高活性突变体构成的基因编辑工具及制备方法和修复先天性视网膜劈裂症致病基因的方法
CN112626070A (zh) * 2021-01-13 2021-04-09 海南微氪生物科技股份有限公司 一种基于切离酶基因编辑技术的定向修复系统
WO2022152746A1 (en) 2021-01-13 2022-07-21 Alia Therapeutics Srl K526d cas9 variants and applications thereof
WO2022238958A1 (en) 2021-05-12 2022-11-17 Crispr Therapeutics Ag Multiplex gene editing
WO2023285431A1 (en) 2021-07-12 2023-01-19 Alia Therapeutics Srl Compositions and methods for allele specific treatment of retinitis pigmentosa
EP4413146A2 (en) * 2021-10-05 2024-08-14 The Board of Trustees of the Leland Stanford Junior University Treatment of polycythemia vera via crispr/aav6 genome editing
CN120265763A (zh) * 2023-10-25 2025-07-04 上海津曼特生物科技有限公司 靶向ttr的基因编辑组合物
CN117887760A (zh) * 2024-01-22 2024-04-16 中国医学科学院北京协和医院 SQSTM1/p62基因新致病突变c.1143delC的敲入小鼠模型及其构建方法
CN118497204B (zh) * 2024-07-16 2024-12-03 西北农林科技大学深圳研究院 一种质粒CRISPR-pCas9n及基因编辑的方法与应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016164797A1 (en) * 2015-04-08 2016-10-13 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Activatable crispr/cas9 for spatial and temporal control of genome editing
US9512446B1 (en) * 2015-08-28 2016-12-06 The General Hospital Corporation Engineered CRISPR-Cas9 nucleases
WO2016205613A1 (en) * 2015-06-18 2016-12-22 The Broad Institute Inc. Crispr enzyme mutations reducing off-target effects

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11685935B2 (en) * 2013-05-29 2023-06-27 Cellectis Compact scaffold of Cas9 in the type II CRISPR system
WO2016123243A1 (en) * 2015-01-28 2016-08-04 The Regents Of The University Of California Methods and compositions for labeling a single-stranded target nucleic acid
EP3265559B1 (en) 2015-03-03 2021-01-06 The General Hospital Corporation Engineered crispr-cas9 nucleases with altered pam specificity
IT201700016321A1 (it) 2017-02-14 2018-08-14 Univ Degli Studi Di Trento Mutanti di cas9 ad alta specificita' e loro applicazioni.

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016164797A1 (en) * 2015-04-08 2016-10-13 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Activatable crispr/cas9 for spatial and temporal control of genome editing
WO2016205613A1 (en) * 2015-06-18 2016-12-22 The Broad Institute Inc. Crispr enzyme mutations reducing off-target effects
US9512446B1 (en) * 2015-08-28 2016-12-06 The General Hospital Corporation Engineered CRISPR-Cas9 nucleases

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BENJAMIN P. KLEINSTIVER, PATTANAYAK VIKRAM, PREW MICHELLE S., TSAI SHENGDAR Q., NGUYEN NHU T., ZHENG ZONGLI, JOUNG J. KEITH: "High-fidelity CRISPR–Cas9 nucleases with no detectable genome-wide off-target effects", NATURE, MACMILLAN JOURNALS LTD., ETC, LONDON, vol. 529, no. 7587, 28 January 2016 (2016-01-28), London, pages 490 - 495, XP055303390, ISSN: 0028-0836, DOI: 10.1038/nature16526 *
CASINI A. ET AL.: "Identification of high-fidelity Cas9 variants using a yeast-based screening", HUMAN GENE THERAPY, vol. 27, no. 11, 18 October 2016 (2016-10-18), 21 October 2016 (2016-10-21), page A41, & CONFERENCE ON CHANGING THE FACE OF MODERN MEDICINE - STEM CELLS AND GENE THERAPY; FLORENCE, ITALY, the whole document *
IAN M. SLAYMAKER, LINYI GAO, BERND ZETSCHE, DAVID A. SCOTT, WINSTON X. YAN, FENG ZHANG: "Rationally engineered Cas9 nucleases with improved specificity", SCIENCE, AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE, US, vol. 351, no. 6268, 1 January 2016 (2016-01-01), US, pages 84 - 88, XP002757561, ISSN: 0036-8075, DOI: 10.1126/science.aad5227 *
M. JINEK, F. JIANG, D. W. TAYLOR, S. H. STERNBERG, E. KAYA, E. MA, C. ANDERS, M. HAUER, K. ZHOU, S. LIN, M. KAPLAN, A. T. IAVARONE: "Structures of Cas9 Endonucleases Reveal RNA-Mediated Conformational Activation", SCIENCE, AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE, vol. 343, no. 6176, 14 March 2014 (2014-03-14), pages 1247997 - 1247997, XP055149157, ISSN: 00368075, DOI: 10.1126/science.1247997 *

Also Published As

Publication number Publication date
IT201700016321A1 (it) 2018-08-14
AU2018221644A1 (en) 2019-08-29
CA3053218A1 (en) 2018-08-23
US20200149020A1 (en) 2020-05-14
EP3583209A1 (en) 2019-12-25
JP2020507347A (ja) 2020-03-12
US11525127B2 (en) 2022-12-13
CN110520528B (zh) 2024-08-13
BR112019016765A2 (pt) 2020-05-26
MX2019009651A (es) 2019-12-18
EP3583209B1 (en) 2024-12-04
EA201991913A1 (ru) 2020-01-22
WO2018149888A1 (en) 2018-08-23
CN110520528A (zh) 2019-11-29
EP3583209C0 (en) 2024-12-04
JP7232531B2 (ja) 2023-03-03
KR20190116407A (ko) 2019-10-14
EP3583209B8 (en) 2025-02-26

Similar Documents

Publication Publication Date Title
JP7232531B2 (ja) 高忠実度ｃａｓ９バリアントおよびそれらの利用
AU2021202581B2 (en) Genome engineering
US12180520B2 (en) Engineered CRISPR-Cas9 nucleases with altered PAM specificity
JP6745391B2 (ja) 遺伝子操作ＣＲＩＳＰＲ−Ｃａｓ９ヌクレアーゼ
TWI716367B (zh) 用於常間回文重複序列叢集（ｃｒｉｓｐｒ）之大量平行組合性基因學
JP2023052051A (ja) Ｖｉ型ｃｒｉｓｐｒオルソログ及び系
CA3111432A1 (en) Novel crispr enzymes and systems
JP2021524272A (ja) ガイドｒｎａ分子および／またはガイドｒｎａ分子／ｒｎａ誘導型ヌクレアーゼ複合体の無痕跡送達のための小胞およびその産生方法
JP2021048882A (ja) Ｒｎａ誘導性ヒトゲノム改変
JP2019514379A (ja) Ｒｎａ誘導型ヌクレアーゼ活性のインビボ高スループット評価のための方法
US20200246488A1 (en) Compositions and methods for treating inflammatory bowel diseases
Fritzen et al. A single aspartate mutation in the conserved catalytic site of Rev3L generates a hypomorphic phenotype in vivo and in vitro
Casini Better safe than sorry: new CRISPR/Cas9 tools for improved genome engineering
Reilly Bypassable essential genes affecting regulation of heterochromatin silencing in S. pombe
Che The roles of histone H3 lysine 56 acetylation in DNA repair
de Leon Ley Developing cellular tools to report oncogene-dependent replication stress
bin Ahmad Farah Construction and initial characterization of yeast strains with dyskeratosis congenita mutations
Ogailan Construction and initial characterization of yeast strains with Dyskeratosis congenita mutations
NZ754904B2 (en) Genome engineering
NZ754904A (en) Genome engineering
NZ754903B2 (en) Genome engineering