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WO2024050544A3 - Enhanced targeted knock-in frequency in host genomes through crispr exonuclease processing - Google Patents

Enhanced targeted knock-in frequency in host genomes through crispr exonuclease processing Download PDF

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Publication number
WO2024050544A3
WO2024050544A3 PCT/US2023/073361 US2023073361W WO2024050544A3 WO 2024050544 A3 WO2024050544 A3 WO 2024050544A3 US 2023073361 W US2023073361 W US 2023073361W WO 2024050544 A3 WO2024050544 A3 WO 2024050544A3
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WO
WIPO (PCT)
Prior art keywords
crispr
frequency
targeted knock
host genomes
enhanced targeted
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/US2023/073361
Other languages
French (fr)
Other versions
WO2024050544A2 (en
Inventor
Jin Li
Hua Yan
Jeffrey W. Habig
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.)
JR Simplot Co
Original Assignee
JR Simplot Co
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 JR Simplot Co filed Critical JR Simplot Co
Publication of WO2024050544A2 publication Critical patent/WO2024050544A2/en
Publication of WO2024050544A3 publication Critical patent/WO2024050544A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
    • C12N15/8213—Targeted insertion of genes into the plant genome by homologous recombination
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPR]
    • 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]

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The present disclosure relates to methods of increasing the efficiency of targeted DNA sequence insertion into host cell genomes using exonucleases and/or silencing of nonhomologous end joining gene(s).
PCT/US2023/073361 2022-09-01 2023-09-01 Enhanced targeted knock-in frequency in host genomes through crispr exonuclease processing Ceased WO2024050544A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202263403200P 2022-09-01 2022-09-01
US63/403,200 2022-09-01

Publications (2)

Publication Number Publication Date
WO2024050544A2 WO2024050544A2 (en) 2024-03-07
WO2024050544A3 true WO2024050544A3 (en) 2024-07-11

Family

ID=88287307

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2023/073361 Ceased WO2024050544A2 (en) 2022-09-01 2023-09-01 Enhanced targeted knock-in frequency in host genomes through crispr exonuclease processing

Country Status (1)

Country Link
WO (1) WO2024050544A2 (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019086460A1 (en) * 2017-10-30 2019-05-09 Kws Saat Se New strategies for precision genome editing
WO2019234132A1 (en) * 2018-06-05 2019-12-12 KWS SAAT SE & Co. KGaA Base editing in polymerase theta deficient plants
WO2022090224A1 (en) * 2020-10-27 2022-05-05 KWS SAAT SE & Co. KGaA Use of enhanced pol theta activity for eukaryotic genome engineering

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013544082A (en) 2010-10-27 2013-12-12 セレクティス Method for increasing the efficiency of double-strand break-induced mutagenesis
EP4062927A1 (en) 2011-02-28 2022-09-28 Seattle Children's Research Institute Coupling endonucleases with end-processing enzymes drives high efficiency gene disruption
EP4234696A3 (en) 2012-12-12 2023-09-06 The Broad Institute Inc. Crispr-cas component systems, methods and compositions for sequence manipulation
US8697359B1 (en) 2012-12-12 2014-04-15 The Broad Institute, Inc. CRISPR-Cas systems and methods for altering expression of gene products
ES2576126T3 (en) 2012-12-12 2016-07-05 The Broad Institute, Inc. Modification by genetic technology and optimization of improved enzyme systems, methods and compositions for sequence manipulation
US10155938B2 (en) 2015-04-14 2018-12-18 City Of Hope Coexpression of CAS9 and TREX2 for targeted mutagenesis

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019086460A1 (en) * 2017-10-30 2019-05-09 Kws Saat Se New strategies for precision genome editing
WO2019234132A1 (en) * 2018-06-05 2019-12-12 KWS SAAT SE & Co. KGaA Base editing in polymerase theta deficient plants
WO2022090224A1 (en) * 2020-10-27 2022-05-05 KWS SAAT SE & Co. KGaA Use of enhanced pol theta activity for eukaryotic genome engineering

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
BUTLER NATHANIEL M. ET AL: "First-generation genome editing in potato using hairy root transformation", PLANT BIOTECHNOLOGY JOURNAL, vol. 18, no. 11, 16 April 2020 (2020-04-16), GB, pages 2201 - 2209, XP093117150, ISSN: 1467-7644, Retrieved from the Internet <URL:https://onlinelibrary.wiley.com/doi/full-xml/10.1111/pbi.13376> DOI: 10.1111/pbi.13376 *
JIA SHOU ET AL: "Precise and Predictable CRISPR Chromosomal Rearrangements Reveal Principles of Cas9-Mediated Nucleotide Insertion", MOLECULAR CELL, vol. 71, no. 4, 19 July 2018 (2018-07-19), AMSTERDAM, NL, pages 498 - 509, XP055765247, ISSN: 1097-2765, DOI: 10.1016/j.molcel.2018.06.021 *
JIA SHOU: "Supplemental Information: Precise and Predictable CRISPR Chromosomal Rearrangements Reveal Principles of Cas9-Mediated Nucleotide Insertion", MOLECULAR CELL, vol. 71, no. 4, 19 July 2018 (2018-07-19), pages 1 - 9, XP093165719, DOI: https://doi.org/10.1016/j.molcel.2018.06.021 *
MIKHAYLOVA E V ET AL: "Available Toolkits for CRISPR/CAS Genome Editing in Plants", RUSSIAN JOURNAL OF PLANT PHYSIOLOGY, MOSCOW, RU, vol. 69, no. 1, 31 January 2022 (2022-01-31), XP037679519, ISSN: 1021-4437, [retrieved on 20220131], DOI: 10.1134/S1021443722010137 *
QIANWEI ZHANG ET AL: "Fusing T5 Exonuclease with Cas9 and Cas12a Increases the Frequency and Size of Deletion at Target Sites", SCIENCE CHINA LIFE SCIENCES, ZHONGGUO KEXUE ZAZHISHE, CHINA, vol. 63, no. 12, 6 May 2020 (2020-05-06), pages 1918 - 1927, XP009548348, ISSN: 1674-7305 *
WEISS TREVOR ET AL: "Optimization of multiplexed CRISPR/Cas9 system for highly efficient genome editing in Setaria viridis", THE PLANT JOURNAL, vol. 104, no. 3, 8 September 2020 (2020-09-08), GB, pages 828 - 838, XP093117157, ISSN: 0960-7412, Retrieved from the Internet <URL:https://onlinelibrary.wiley.com/doi/full-xml/10.1111/tpj.14949> DOI: 10.1111/tpj.14949 *

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