Harms et al have created this detailed set of protocols for conducting CRISPR/Cas9 mutagenesis and editing in mouse embryos. Included are more guidelines and instructions for choosing targets, then on to ligation protocols for cloning protospacers in sgRNA expression vectors, in vitro RNA synthesis, pronuclear injection, and follow-up screening/genotyping of founder animals, as well as HDR donor design and considerations. Also a timeline schematic.
Mouse Genome Editing Using the CRISPR/Cas System.Harms DW, Quadros RM, Seruggia D, Ohtsuka M, Takahashi G, Montoliu L, Gurumurthy CB.Curr Protoc Hum Genet. 2014 Oct 1;83:15.7.1-15.7.27. doi: 10.1002/0471142905.hg1507s83.
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