Mouse Genome Editing Using the CRISPR/Cas System

Curr Protoc Hum Genet. 2014 Oct 1:83:15.7.1-27. doi: 10.1002/0471142905.hg1507s83.

Abstract

The availability of techniques to create desired genetic mutations has enabled the laboratory mouse as an extensively used model organism in biomedical research including human genetics. A new addition to this existing technical repertoire is the CRISPR/Cas system. Specifically, this system allows editing of the mouse genome much more quickly than the previously used techniques, and, more importantly, multiple mutations can be created in a single experiment. Here we provide protocols for preparation of CRISPR/Cas reagents and microinjection into one-cell mouse embryos to create knockout or knock-in mouse models.

Keywords: CRISPR/Cas9; gene editing; mutant mouse; pronuclear and cytoplasmic injection; sgRNA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Genome*
  • Mice