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

Autor: Dawn E. Watkins-Chow, Gaurav K. Varshney, Lisa J. Garrett, Zelin Chen, Erin A. Jimenez, Cecilia Rivas, Kevin S. Bishop, Raman Sood, Ursula L. Harper, William J. Pavan, Shawn M. Burgess
Jazyk: angličtina
Rok vydání: 2017
Předmět:
Zdroj: G3: Genes, Genomes, Genetics, Vol 7, Iss 2, Pp 719-722 (2017)
Druh dokumentu: article
ISSN: 2160-1836
DOI: 10.1534/g3.116.038091
Popis: 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.
Databáze: Directory of Open Access Journals