Highly-efficient Cpf1-mediated gene targeting in mice following high concentration pronuclear injection
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 co-injecting two guides with close targeting sites can result in synergistic genomic cutting, even if one of the guides has minimal cutting activity.
Watkins-Chow Dawn E.、Varshney Gaurav K.、Chen Zelin、Bishop Kevin S.、Harper Ursula L.、Garrett Lisa J.、Rivas Cecilia、Pavan William J.、Sood Raman、Burgess Shawn M.、Jimenez Erin A.
Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of HealthFunctional and Chemical Genomics Program, Oklahoma Medical Research FoundationTranslational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of HealthZebrafish Core, Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of HealthGenomics Core, National Human Genome Research Institute, National Institutes of HealthEmbryonic Stem Cell and Transgenic Mouse Core, National Human Genome Research Institute, National Institutes of HealthEmbryonic Stem Cell and Transgenic Mouse Core, National Human Genome Research Institute, National Institutes of HealthGenetic Disease Research Branch, National Human Genome Research Institute, National Institutes of HealthZebrafish Core, Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of HealthTranslational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of HealthTranslational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health
遗传学分子生物学生物工程学
Cpf1 nucleasegenome editingCRISPRmouse
Watkins-Chow Dawn E.,Varshney Gaurav K.,Chen Zelin,Bishop Kevin S.,Harper Ursula L.,Garrett Lisa J.,Rivas Cecilia,Pavan William J.,Sood Raman,Burgess Shawn M.,Jimenez Erin A..Highly-efficient Cpf1-mediated gene targeting in mice following high concentration pronuclear injection[EB/OL].(2025-03-28)[2025-05-04].https://www.biorxiv.org/content/10.1101/097220.点此复制
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