Editing the Mouse Genome Using the CRISPR-Cas9 System

Cold Spring Harb Protoc. 2016 Feb 1;2016(2):pdb.top087536. doi: 10.1101/pdb.top087536.

Abstract

The ability to modify the murine genome is perhaps one of the most important developments in modern biology. However, traditional methods of genomic engineering are costly and relatively clumsy in their approach. The use of programmable nucleases such as zinc finger nucleases and transcription activator-like effector nucleases significantly improved the precision of genome-editing technology, but the design and use of these nucleases remains cumbersome and prohibitively expensive. The CRISPR-Cas9 system is the next installment in the line of programmable nucleases; it provides highly efficient and precise genome-editing capabilities using reagents that are simple to design and inexpensive to generate. Furthermore, with the CRISPR-Cas9 system, it is possible to move from a hypothesis to an in vivo mouse model in less than a month. The simplicity, cost effectiveness, and speed of the CRISPR-Cas9 system allows researchers to tackle questions that otherwise would not be technically or financially viable. In this introduction, we discuss practical considerations for the use of Cas9 in genome engineering in mice.

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Gene Targeting / methods*
  • Genome*
  • Mice
  • Recombination, Genetic*