Recent Advances in Improving Gene-Editing Specificity through CRISPR-Cas9 Nuclease Engineering

Cells. 2022 Jul 13;11(14):2186. doi: 10.3390/cells11142186.

Abstract

CRISPR-Cas9 is the state-of-the-art programmable genome-editing tool widely used in many areas. For safe therapeutic applications in clinical medicine, its off-target effect must be dramatically minimized. In recent years, extensive studies have been conducted to improve the gene-editing specificity of the most popular CRISPR-Cas9 nucleases using different strategies. In this review, we summarize and discuss these strategies and achievements, with a major focus on improving the gene-editing specificity through Cas9 protein engineering.

Keywords: CRISPR–Cas9; gene editing; high fidelity; off-target; protein engineering; specificity.

Publication types

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

MeSH terms

  • CRISPR-Associated Protein 9 / genetics
  • CRISPR-Cas Systems* / genetics
  • Endonucleases / genetics
  • Gene Editing*
  • Protein Engineering

Substances

  • CRISPR-Associated Protein 9
  • Endonucleases