Generation of site-specific mutations in the rat genome via CRISPR/Cas9

Methods Enzymol. 2014;546:297-317. doi: 10.1016/B978-0-12-801185-0.00014-3.

Abstract

The laboratory rat is a valuable model organism for basic biological studies and drug development. However, due to the lack of genetic tools for site-specific genetic modification in the rat genome, more and more researchers chose the mouse as their favored mammalian models due to the sophisticated embryonic stem cell-based gene-targeting techniques available. Recently, engineered nucleases, including zinc finger nucleases, transcription activator-like effector nucleases, and CRISPR/Cas9 systems, have been adapted to generate knockout rats efficiently. The purpose of this section is to provide detailed procedures for the generation of site-specific mutations in the rat genome through injection of Cas9/sgRNA into one-cell embryos.

Keywords: Gene editing; Knockin; Knockout; Mutation; One-cell embryo; Rat genome.

MeSH terms

  • Animals
  • CRISPR-Associated Proteins / genetics
  • CRISPR-Cas Systems
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Embryo, Mammalian / cytology*
  • Embryo, Mammalian / metabolism
  • Endonucleases / genetics
  • Female
  • Gene Targeting / methods*
  • Genome
  • Mutagenesis*
  • Mutation
  • Rats

Substances

  • CRISPR-Associated Proteins
  • Endonucleases