Genome Editing in Mouse and Rat by Electroporation

Methods Mol Biol. 2017;1630:81-89. doi: 10.1007/978-1-4939-7128-2_7.

Abstract

Many knock-out/knock-in mouse and rat strains have been produced by genome editing techniques using engineered endonucleases, including zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), or clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9. Microinjection of engineered endonucleases into pronuclear-stage embryos is required to produce genome-edited rodents and the development of easy, rapid, and high-efficiency methods that do not require special skills such as microinjection is needed. This chapter presents a new technique called Technique for Animal Knockout system by Electroporation (TAKE), which produces genome-edited rodents by direct introduction of engineered endonucleases into intact embryos using electroporation.

Keywords: CRISPR/Cas9; Cas9 protein; Electroporation; Intact embryos; Knock-in; Knock-out; Mouse; Rat; TALEN; ZFN.

MeSH terms

  • Animals
  • CRISPR-Associated Proteins / metabolism
  • Electroporation / methods*
  • Endonucleases / metabolism*
  • Female
  • Gene Editing / methods*
  • Male
  • Mice
  • Mice, Knockout
  • RNA, Guide / genetics
  • Rats
  • Transcription Activator-Like Effector Nucleases / metabolism
  • Zinc Finger Nucleases / metabolism

Substances

  • CRISPR-Associated Proteins
  • RNA, Guide
  • Endonucleases
  • Transcription Activator-Like Effector Nucleases
  • Zinc Finger Nucleases