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

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 gu...

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Detalles Bibliográficos
Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Genetics Society of America 2016
Materias:
Investigations
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5295614/
https://www.ncbi.nlm.nih.gov/pubmed/28040780
http://dx.doi.org/10.1534/g3.116.038091
Descripción
Sumario: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.

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