Introduction to Gene Editing and Manipulation Using CRISPR/Cas9 Technology

Curr Protoc Mol Biol. 2016 Jul 1:115:31.4.1-31.4.6. doi: 10.1002/cpmb.14.

Abstract

Until very recently, the prospect of introducing mutations or exogenous DNA sequences at precise locations in the genomes of plants and animals was difficult, if not impossible. This rapidly changed with the demonstration that the type II CRISPR-Cas complex, a bacterial anti-viral surveillance system, could be engineered into a simple and robust platform for introducing double-stranded DNA breaks at nearly any position of plant and animal genomes. The prospect of efficiently creating tailored changes to a gene of interest is revolutionizing biomedical research, allowing exciting new questions to be asked. This overview introduces CRISPR-Cas technology as a tool for molecular biology and briefly discusses the advantages of this method over earlier techniques, as well as unique opportunities to create new avenues of research. © 2016 by John Wiley & Sons, Inc.

Keywords: CRISPR; Cas9; genome editing; sgRNA.

Publication types

  • Review

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • CRISPR-Associated Protein 9
  • CRISPR-Cas Systems*
  • DNA Breaks, Double-Stranded
  • Endonucleases / genetics
  • Gene Editing / methods*
  • Genome
  • Humans
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • Streptococcus pyogenes / genetics

Substances

  • Bacterial Proteins
  • RNA, Guide, CRISPR-Cas Systems
  • CRISPR-Associated Protein 9
  • Cas9 endonuclease Streptococcus pyogenes
  • Endonucleases