Improving CRISPR Genome Editing by Engineering Guide RNAs

Trends Biotechnol. 2019 Aug;37(8):870-881. doi: 10.1016/j.tibtech.2019.01.009. Epub 2019 Mar 4.

Abstract

CRISPR technology is a two-component gene editing system in which the effector protein induces genetic alterations with the aid of a gene targeting guide RNA. Guide RNA can be produced through chemical synthesis, in vitro transcription, or intracellular transcription. Guide RNAs can be engineered to have chemical modifications, alterations in the spacer length, sequence modifications, fusion of RNA or DNA components, and incorporation of deoxynucleotides. Engineered guide RNA can improve genome editing efficiency and target specificity, regulation of biological toxicity, sensitive and specific molecular imaging, multiplexing, and editing flexibility. Therefore, engineered guide RNA will enable more specific, efficient, and safe gene editing, ultimately improving the clinical benefits of gene therapy.

Keywords: CRISPR; gene therapy; genome editing; guide RNA; guide RNA engineering.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Endoribonucleases / metabolism*
  • Gene Editing / methods*
  • Gene Editing / trends
  • Genetic Engineering / methods*
  • Genetic Engineering / trends
  • RNA, Guide, CRISPR-Cas Systems / genetics*
  • RNA, Guide, CRISPR-Cas Systems / metabolism
  • Substrate Specificity

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • Endoribonucleases