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 coinjecti...

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Bibliographic Details
Published in:G3 : genes - genomes - genetics 2017-02, Vol.7 (2), p.719-722
Main Authors: Watkins-Chow, Dawn E, Varshney, Gaurav K, Garrett, Lisa J, Chen, Zelin, Jimenez, Erin A, Rivas, Cecilia, Bishop, Kevin S, Sood, Raman, Harper, Ursula L, Pavan, William J, Burgess, Shawn M
Format: Article
Language:English
Subjects:
Acidaminococcus - enzymology
Acidaminococcus - genetics
Animals
Bacterial Proteins - genetics
Cpf1 nuclease
CRISPR
CRISPR-Cas Systems - genetics
Endonucleases - genetics
Gene Editing - methods
Gene Targeting - methods
genome editing
Investigations
Mice
mouse
RNA, Guide, CRISPR-Cas Systems - genetics
RNA, Messenger - genetics
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Summary: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.
ISSN:2160-1836
2160-1836
DOI:10.1534/g3.116.038091