Improving CRISPR-Cas9 On-Target Specificity

Curr Issues Mol Biol. 2018:26:65-80. doi: 10.21775/cimb.026.065. Epub 2017 Sep 7.

Abstract

The CRISPR-Cas9 has revolutionized the field of molecular biology, medical genetics and medicine. The technology is robust, facile and simple to achieve genome targeting in cells and organisms. However, to propagate these nucleases for therapeutic application, the on-target specificity is of paramount importance. Although the binding and cleavage of off-target sites by Cas9 is issue of concern, however the specificity of CRISPR technology is greatly improved in current research employing the use of engineer nucleases, improved gRNA selection, novel Cas9 orhtologs and the advancement in methods to detect and screen off-target sites and its effects. Here we summarize the advances in this state-of-the-art technology that will equip the genome editing tools to be applied in clinical research. The researcher should optimize these methods with emphasize to achieve perfection in the specificity.

Publication types

  • Review

MeSH terms

  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • CRISPR-Associated Protein 9
  • CRISPR-Cas Systems*
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Endonucleases / genetics*
  • Endonucleases / metabolism
  • Gene Targeting
  • Genetic Engineering / methods*
  • Genome, Human*
  • Humans
  • Internet
  • Mutation
  • Plasmids / chemistry
  • Plasmids / metabolism
  • RNA, Guide, CRISPR-Cas Systems / genetics*
  • RNA, Guide, CRISPR-Cas Systems / metabolism
  • SELEX Aptamer Technique
  • Sensitivity and Specificity
  • Software

Substances

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