Title

Efficient CRISPR/Cas9-Mediated Genome Editing in Mice by Zygote Electroporation of Nuclease.

Document Type

Article

Publication Date

6-2015

JAX Source

Genetics 2015 Jun; 200:423-430.

PMID

25819794

Grant

CA034196, DK097605

Abstract

CRISPR/Cas is an adaptive immune system in bacteria and archaea that has recently been exploited for genome engineering. Mutant mice can be generated in one step through direct delivery of the CRISPR/Cas9 components into a mouse zygote. Although the technology is robust, delivery remains a bottleneck, as it involves manual injection of the components into the pronuclei or the cytoplasm of mouse zygotes, which is technically demanding and inherently low throughput. To overcome this limitation, we employed electroporation as a means to deliver the CRISPR/Cas9 components, including Cas9 mRNA, sgRNA, and donor oligonucleotide, into mouse zygotes and recovered live mice with targeted NHEJ and HDR mutations with high efficiency. Our results demonstrate that mice carrying CRISPR/Cas9-mediated targeted mutations can be obtained with high efficiency by zygote electroporation. Genetics 2015 Jun; 200:423-430.