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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.

SUBMITTER: Watkins-Chow DE

PROVIDER: S-EPMC5295614 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature

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Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection.

Watkins-Chow Dawn E DE Varshney Gaurav K GK Garrett Lisa J LJ Chen Zelin Z Jimenez Erin A EA Rivas Cecilia C Bishop Kevin S KS Sood Raman R Harper Ursula L UL Pavan William J WJ Burgess Shawn M SM

G3 (Bethesda, Md.) 20170209 2

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. ...[more]

PMID: 28040780

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