Mouse genome engineering via CRISPR-Cas9 for study of immune function

Immunity. 2015 Jan 20;42(1):18-27. doi: 10.1016/j.immuni.2015.01.004.

Abstract

Clustered regularly interspaced palindromic repeats (CRISPR)-associated (Cas9) technology has proven a formidable addition to our armory of approaches for genomic editing. Derived from pathways in archaea and bacteria that mediate the resistance to exogenous genomic material, the CRISPR-Cas9 system utilizes a short single guide RNA (sgRNA) to direct the endonuclease Cas9 to virtually anywhere in the genome. Upon targeting, Cas9 generates DNA double-strand breaks (DSBs) and facilitates the repair or insertion of mutations, insertion of recombinase recognition sites, or large DNA elements. Here, we discuss the practical advantages of the CRISPR-Cas9 system over conventional and other nuclease-based targeting technologies and provide suggestions for the use of this technology to address immunological questions.

Publication types

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

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics*
  • DNA Repair
  • Endonucleases / metabolism
  • Genetic Engineering / methods*
  • Genome* / genetics
  • Humans
  • Immunity / genetics*
  • Mice
  • RNA, Guide / genetics

Substances

  • RNA, Guide
  • Endonucleases