Gene editing in mouse zygotes using the CRISPR/Cas9 system
Methods. 2017 Mar 2. pii: S1046-2023(16)30469-8. doi: 10.1016/j.ymeth.2017.02.008. [Epub ahead of print]
|Authors/Editors:||Wefers B, Bashir S, Rossius J, Wurst W, Kühn R.|
The generation of targeted mouse mutants is a key technology for biomedical research. Using the CRISPR/Cas9 system for induction of targeted double-strand breaks, gene editing can be performed in a single step directly in mouse zygotes. This article covers the design of knockout and knockin alleles, preparation of reagents, microinjection or electroporation of zygotes and the genotyping of pups derived from gene editing projects. In addition we include a section for the control of experimental settings by targeting the Rosa26 locus and PCR based genotyping of blastocysts.