WO2019023291A3 - Compositions and methods for making and decoding paired-guide rna libraries and uses thereof - Google Patents

Compositions and methods for making and decoding paired-guide rna libraries and uses thereof Download PDF

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Publication number
WO2019023291A3
WO2019023291A3 PCT/US2018/043588 US2018043588W WO2019023291A3 WO 2019023291 A3 WO2019023291 A3 WO 2019023291A3 US 2018043588 W US2018043588 W US 2018043588W WO 2019023291 A3 WO2019023291 A3 WO 2019023291A3
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Prior art keywords
paired
decoding
compositions
making
methods
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PCT/US2018/043588
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French (fr)
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WO2019023291A2 (en
Inventor
Xiaole LIU
Myles Brown
Jingyu PENG
Tengfei XIAO
Wei Li
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Dana-Farber Cancer Institute, Inc.
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Publication of WO2019023291A2 publication Critical patent/WO2019023291A2/en
Publication of WO2019023291A3 publication Critical patent/WO2019023291A3/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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/111—General methods applicable to biologically active non-coding nucleic acids
    • 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 [CRISPRs]
    • 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
    • C12N2320/00—Applications; Uses
    • C12N2320/10—Applications; Uses in screening processes
    • C12N2320/12—Applications; Uses in screening processes in functional genomics, i.e. for the determination of gene function
    • 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
    • C12N2330/00—Production
    • C12N2330/30—Production chemically synthesised
    • C12N2330/31—Libraries, arrays

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

Abstract

The present disclosure relates to compositions and methods for making and decoding paired-guide RNA (pgRNA) libraries using the Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR) system, and using the resulting pgRNA/CRISPR libraries to identify genetic interactions or functional non-coding elements.
PCT/US2018/043588 2017-07-25 2018-07-25 Compositions and methods for making and decoding paired-guide rna libraries and uses thereof WO2019023291A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201762536870P 2017-07-25 2017-07-25
US62/536,870 2017-07-25

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WO2019023291A2 WO2019023291A2 (en) 2019-01-31
WO2019023291A3 true WO2019023291A3 (en) 2019-04-25

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EP2841572B1 (en) 2012-04-27 2019-06-19 Duke University Genetic correction of mutated genes
EP3362571A4 (en) 2015-10-13 2019-07-10 Duke University Genome engineering with type i crispr systems in eukaryotic cells
US20210017516A1 (en) * 2019-07-16 2021-01-21 Massachusetts Institute Of Technology Methods of multiplexing crispr
US20230201375A1 (en) * 2020-04-27 2023-06-29 Duke University Targeted genomic integration to restore neurofibromin coding sequence in neurofibromatosis type 1 (nf1)
EP4288551A1 (en) * 2021-02-02 2023-12-13 Limagrain Europe Linkage of a distal promoter to a gene of interest by gene editing to modify gene expression

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US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
DK2898075T3 (en) 2012-12-12 2016-06-27 Broad Inst Inc CONSTRUCTION AND OPTIMIZATION OF IMPROVED SYSTEMS, PROCEDURES AND ENZYME COMPOSITIONS FOR SEQUENCE MANIPULATION
CN114634950A (en) 2012-12-12 2022-06-17 布罗德研究所有限公司 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
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US20150225730A1 (en) * 2014-02-12 2015-08-13 Dna2.0, Inc. Methods for generating libraries with co-varying regions of polynuleotides for genome modification
WO2016130697A1 (en) * 2015-02-11 2016-08-18 Memorial Sloan Kettering Cancer Center Methods and kits for generating vectors that co-express multiple target molecules
WO2017069829A2 (en) * 2015-07-31 2017-04-27 The Trustees Of Columbia University In The City Of New York High-throughput strategy for dissecting mammalian genetic interactions
WO2017122096A1 (en) * 2016-01-15 2017-07-20 Astrazeneca Ab Gene modification assays

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CARLOS PULIDO-QUETGLAS ET AL: "Scalable Design of Paired CRISPR Guide RNAs for Genomic Deletion", PLOS COMPUTATIONAL BIOLOGY, vol. 13, no. 3, 2 March 2017 (2017-03-02), pages e1005341, XP055545792, DOI: 10.1371/journal.pcbi.1005341 *
DOW LUKAS E ET AL: "Inducible in vivo genome editing with CRISPR-Cas9", NATURE BIOTECHNOLOGY, vol. 33, no. 4, 18 February 2015 (2015-02-18), pages 390, XP055266912 *
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