Diverse Class 2 CRISPR Effectors as Active Nucleases with Expanded Targeting Capabilities

CRISPR J. 2024 Apr;7(2):120-130. doi: 10.1089/crispr.2023.0058.

Abstract

CRISPR-Cas systems have proven effective in a variety of applications due to their ease of use and relatively high editing efficiency. Yet, any individual CRISPR-Cas system has inherent limitations, necessitating a diversity of RNA-guided nucleases to suit applications with distinct needs. We searched through metagenomic sequences to identify RNA-guided nucleases and found enzymes from diverse CRISPR-Cas types and subtypes, the most promising of which we developed into gene-editing platforms. Based on prior annotations of the metagenomic sequences, we establish the likely taxa and sampling locations where Class 2 CRISPR-Cas systems active in eukaryotes may be found. The newly discovered systems show robust capabilities as gene editors and base editors.

Publication types

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

MeSH terms

  • CRISPR-Cas Systems* / genetics
  • Endonucleases / genetics
  • Gene Editing*
  • RNA

Substances

  • Endonucleases
  • RNA