Mechanistic insights into the versatile class II CRISPR toolbox

Fan Zhang; Zhiwei Huang

doi:10.1016/j.tibs.2021.11.007

Mechanistic insights into the versatile class II CRISPR toolbox

Trends Biochem Sci. 2022 May;47(5):433-450. doi: 10.1016/j.tibs.2021.11.007. Epub 2021 Dec 14.

Authors

Fan Zhang¹, Zhiwei Huang²

Affiliations

¹ HIT Center for Life Sciences, School of Life Science and Technology, Harbin Institute of Technology, Harbin 150080, China.
² HIT Center for Life Sciences, School of Life Science and Technology, Harbin Institute of Technology, Harbin 150080, China. Electronic address: huangzhiwei@hit.edu.cn.

PMID: 34920928
DOI: 10.1016/j.tibs.2021.11.007

Abstract

The constantly expanding group of class II CRISPR-Cas (clustered regularly interspaced short palindromic repeats-associated) effectors and their engineered variants exhibit distinct editing modes and efficiency, fidelity, target range, and molecular size. Their enormous diversity of capabilities provides a formidable toolkit for a large array of technologies. We review the structural and biochemical mechanisms of versatile effector proteins from class II CRISPR-Cas systems to provide mechanistic insights into their target specificity, protospacer adjacent motif (PAM) restriction, and activity regulation, and discuss possible strategies to enhance genome-engineering tools in terms of accuracy, efficiency, applicability, and controllability.

Keywords: CRISPR-Cas; PAM restriction; anti-CRISPR; rational engineering; structural mechanism; target specificity.

Publication types

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

MeSH terms

CRISPR-Cas Systems* / genetics
Gene Editing*