title: Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection.
abstract: Cpf1 has emerged as an alternative to the Cas9 RNA-guided nuclease. Here we show that gene targeting rates in mice using Cpf1 can meet, or even surpass, Cas9 targeting rates (approaching 100% targeting), but require higher concentrations of mRNA and guide. We also demonstrate that coinjecting two guides with close targeting sites can result in synergistic genomic cutting, even if one of the guides has minimal cutting activity.
authors.affiliation: Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Dawn E
authors.identifier: 0000-0002-4355-0868
authors.initials: DE
authors.lastname: Watkins-Chow
authors.affiliation: Functional and Chemical Genomics Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104.
authors.forename: Gaurav K
authors.identifier: 0000-0002-0429-1904
authors.initials: GK
authors.lastname: Varshney
authors.affiliation: Embryonic Stem Cell and Transgenic Mouse Core, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Garrett
authors.affiliation: Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Zelin
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Erin A
authors.initials: EA
authors.lastname: Jimenez
authors.affiliation: Embryonic Stem Cell and Transgenic Mouse Core, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Rivas
authors.affiliation: Zebrafish Core, Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Kevin S
authors.initials: KS
authors.lastname: Bishop
authors.affiliation: Zebrafish Core, Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Raman
authors.identifier: 0000-0001-5565-662X
authors.initials: R
authors.lastname: Sood
authors.affiliation: Genomics Core, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Ursula L
authors.initials: UL
authors.lastname: Harper
authors.affiliation: Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892 bpavan@mail.nih.gov burgess@mail.nih.gov.
authors.forename: William J
authors.identifier: 0000-0001-8281-5120
authors.initials: WJ
authors.lastname: Pavan
authors.affiliation: Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892 bpavan@mail.nih.gov burgess@mail.nih.gov.
authors.forename: Shawn M
authors.identifier: 0000-0003-1147-0596
authors.initials: SM
authors.lastname: Burgess
mesh_terms: D045850:Acidaminococcus / Q000201:enzymology / Q000235:genetics; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D064113:CRISPR-Cas Systems / Q000235:genetics; D004720:Endonucleases / Q000235:genetics*; D000072669:Gene Editing / Q000379:methods*; D018390:Gene Targeting / Q000379:methods*; D051379:Mice; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D004706:Endodeoxyribonucleases; D064130:CRISPR-Associated Proteins
chemical_list: D001426:Bacterial Proteins; D004720:Endonucleases; D000094704:RNA, Guide, CRISPR-Cas Systems; D012333:RNA, Messenger; C000656745:Cas12a protein; D004706:Endodeoxyribonucleases; D064130:CRISPR-Associated Proteins
keywords: CRISPR; Cpf1 nuclease; genome editing; mouse