CA3084020C - Modified CAS9 systems to alter the genome of eukaryotes - Google Patents

Modified CAS9 systems to alter the genome of eukaryotes

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Publication number
CA3084020C
CA3084020C CA3084020A CA3084020A CA3084020C CA 3084020 C CA3084020 C CA 3084020C CA 3084020 A CA3084020 A CA 3084020A CA 3084020 A CA3084020 A CA 3084020A CA 3084020 C CA3084020 C CA 3084020C
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Prior art keywords
sequence
engineered
protein
cell
cas9 protein
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CA3084020A
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English (en)
French (fr)
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CA3084020A1 (en
Inventor
Timothy Seebeck
Fuqiang Chen
Gregory Davis
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Sigma Aldrich Co LLC
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/87Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90Stable introduction of foreign DNA into chromosome
    • C12N15/902Stable introduction of foreign DNA into chromosome using homologous recombination
    • C12N15/907Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/102Mutagenizing nucleic acids
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/11DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/16Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • C07K2319/01Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/09Fusion polypeptide containing a localisation/targetting motif containing a nuclear localisation signal
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • C07K2319/01Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/10Fusion polypeptide containing a localisation/targetting motif containing a tag for extracellular membrane crossing, e.g. TAT or VP22
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • C07K2319/85Fusion polypeptide containing an RNA binding domain
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/10Type of nucleic acid
    • C12N2310/20Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPR]
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y301/00Hydrolases acting on ester bonds (3.1)
    • C12Y301/21Endodeoxyribonucleases producing 5'-phosphomonoesters (3.1.21)
    • C12Y301/21001Deoxyribonuclease I (3.1.21.1)

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Cell Biology (AREA)
  • Mycology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
CA3084020A 2018-02-15 2019-02-15 Modified CAS9 systems to alter the genome of eukaryotes Active CA3084020C (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US201862631304P 2018-02-15 2018-02-15
US62/631,304 2018-02-15
US201862720525P 2018-08-21 2018-08-21
US62/720,525 2018-08-21
PCT/US2019/018335 WO2019161290A1 (en) 2018-02-15 2019-02-15 Engineered cas9 systems for eukaryotic genome modification

Related Child Applications (1)

Application Number Title Priority Date Filing Date
CA3281020 Division 2019-02-15

Publications (2)

Publication Number Publication Date
CA3084020A1 CA3084020A1 (en) 2019-08-22
CA3084020C true CA3084020C (en) 2025-11-25

Family

ID=65995829

Family Applications (1)

Application Number Title Priority Date Filing Date
CA3084020A Active CA3084020C (en) 2018-02-15 2019-02-15 Modified CAS9 systems to alter the genome of eukaryotes

Country Status (11)

Country Link
US (2) US10767193B2 (enExample)
EP (1) EP3752607A1 (enExample)
JP (3) JP7109547B2 (enExample)
KR (3) KR102684890B1 (enExample)
CN (1) CN111902536B (enExample)
AU (3) AU2019222568B2 (enExample)
BR (1) BR112020010479A2 (enExample)
CA (1) CA3084020C (enExample)
IL (2) IL274528B2 (enExample)
SG (1) SG11202007382TA (enExample)
WO (1) WO2019161290A1 (enExample)

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EP3204496A1 (en) 2014-10-10 2017-08-16 Editas Medicine, Inc. Compositions and methods for promoting homology directed repair
US11111492B2 (en) * 2017-03-06 2021-09-07 Florida State University Research Foundation, Inc. Genome engineering methods using a cytosine-specific Cas9
WO2019222545A1 (en) 2018-05-16 2019-11-21 Synthego Corporation Methods and systems for guide rna design and use
CA3104856A1 (en) 2018-06-29 2020-01-02 Editas Medicine, Inc. Synthetic guide molecules, compositions and methods relating thereto
US10982200B2 (en) 2019-02-14 2021-04-20 Metagenomi Ip Technologies, Llc Enzymes with RuvC domains
JP2022520104A (ja) 2019-02-15 2022-03-28 シグマ-アルドリッチ・カンパニー・リミテッド・ライアビリティ・カンパニー Crispr/cas融合タンパク質およびシステム
WO2020181101A1 (en) 2019-03-07 2020-09-10 The Regents Of The University Of California Crispr-cas effector polypeptides and methods of use thereof
WO2021127209A1 (en) 2019-12-17 2021-06-24 Sigma-Aldrich Co. Llc Genome editing in bacteroides
GB2594339B (en) * 2020-02-14 2023-02-08 Metagenomi Inc Enzymes with RUVC domains
BR112022012350A2 (pt) * 2020-03-11 2022-09-27 Sigma Aldrich Co Llc Nucleases de spcas9 de alta fidelidade para modificação de genoma
MX2022012110A (es) * 2020-03-31 2022-10-18 Metagenomi Inc Sistemas crispr clase ii tipo ii.
CN112159801B (zh) * 2020-07-13 2022-11-15 复旦大学 SlugCas9-HF蛋白、含有SlugCas9-HF蛋白的基因编辑系统及应用
EP4230737A4 (en) * 2020-09-21 2024-07-24 Korea University Research and Business Foundation NOVEL IMPROVED BASIC EDITING OR REVISION FUSION PROTEIN AND USE THEREOF
MX2023005150A (es) * 2020-11-06 2023-05-26 Pairwise Plants Services Inc Composiciones y metodos para el reemplazo de alelos con adn codificado por arn.
WO2023028348A1 (en) 2021-08-27 2023-03-02 Metagenomi, Inc. Enzymes with ruvc domains
CN113717960B (zh) * 2021-08-27 2023-07-18 电子科技大学 一种新Cas9蛋白、CRISPR-Cas9基因组定向编辑载体及基因组编辑方法
WO2023097282A1 (en) 2021-11-24 2023-06-01 Metagenomi, Inc. Endonuclease systems
CN114657181B (zh) * 2022-04-01 2023-08-25 安徽大学 一种靶向H1.4的sgRNA以及H1.4基因编辑方法

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CA2468292A1 (en) 2001-11-26 2003-06-05 Advanced Cell Technology, Inc. Methods for making and using reprogrammed human somatic cell nuclei and autologous and isogenic human stem cells
GB2537000C (en) 2012-05-25 2019-10-09 Univ California Methods and compositions for RNA-directed target DNA modification and for RNA-directed modulation of transcription
CA2891347C (en) * 2012-12-06 2018-02-27 Sigma-Aldrich Co. Llc Crispr-based genome modification and regulation
KR20150105635A (ko) * 2012-12-12 2015-09-17 더 브로드 인스티튜트, 인코퍼레이티드 서열 조작을 위한 crispr-cas 성분 시스템, 방법 및 조성물
EP4435110A3 (en) 2013-06-05 2025-01-01 Duke University Rna-guided gene editing and gene regulation
US9074199B1 (en) * 2013-11-19 2015-07-07 President And Fellows Of Harvard College Mutant Cas9 proteins
EP3097212A4 (en) * 2014-01-24 2017-10-04 North Carolina State University Methods and compositions for sequences guiding cas9 targeting
US11242525B2 (en) 2014-03-26 2022-02-08 Editas Medicine, Inc. CRISPR/CAS-related methods and compositions for treating sickle cell disease
WO2016033298A1 (en) * 2014-08-28 2016-03-03 North Carolina State University Novel cas9 proteins and guiding features for dna targeting and genome editing
EP3845655B1 (en) * 2014-10-01 2025-01-01 The General Hospital Corporation Methods for increasing efficiency of nuclease-induced homology-directed repair
PL3272867T3 (pl) * 2016-06-02 2020-01-31 Sigma-Aldrich Co. Llc Wykorzystanie programowalnych białek wiążących dna w celu wzmocnienia celowanej modyfikacji genomu
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CN111630170B (zh) * 2017-07-31 2025-03-07 映像生物有限公司 眼部疾病的细胞模型及用于眼部疾病的疗法

Also Published As

Publication number Publication date
BR112020010479A2 (pt) 2020-11-24
US10767193B2 (en) 2020-09-08
US20200354752A1 (en) 2020-11-12
JP2022122910A (ja) 2022-08-23
KR102494449B1 (ko) 2023-01-31
AU2024202158A1 (en) 2024-05-02
US12297449B2 (en) 2025-05-13
JP2021505180A (ja) 2021-02-18
IL274528A (en) 2020-06-30
KR102684890B1 (ko) 2024-07-12
AU2019222568B2 (en) 2021-10-14
EP3752607A1 (en) 2020-12-23
US20190249200A1 (en) 2019-08-15
KR20230022258A (ko) 2023-02-14
JP7109547B2 (ja) 2022-07-29
AU2022200130B2 (en) 2024-03-07
CN111902536B (zh) 2024-07-30
AU2019222568A1 (en) 2020-05-21
KR20220015502A (ko) 2022-02-08
JP2024161376A (ja) 2024-11-19
WO2019161290A1 (en) 2019-08-22
KR102465067B1 (ko) 2022-11-10
AU2022200130A1 (en) 2022-02-10
IL321024A (en) 2025-07-01
JP7559005B2 (ja) 2024-10-01
SG11202007382TA (en) 2020-08-28
IL274528B2 (en) 2025-10-01
IL274528B1 (en) 2025-06-01
KR20200121342A (ko) 2020-10-23
CN111902536A (zh) 2020-11-06
CA3084020A1 (en) 2019-08-22

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