基于CRISPR/Cas系统的多重基因编辑与调控技术

Multiplex gene editing and regulation techniques based on CRISPR/Cas system

规律成簇的间隔短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)及其相关Cas蛋白所构建的CRISPR/Cas系统是古细菌或细菌中特有的一种获得性免疫系统。研究人员将其开发成基因编辑工具之后,凭借其高效、精准和通用性强等优点迅速成为合成生物学领域的热门研究方向,在生命科学、生物工程技术、食品科学及农作物育种等多个领域引发了革命性的影响。目前基于CRISPR/Cas系统单基因编辑与调控技术日益完善,但在多重基因编辑和调控方面仍存在挑战。本文聚焦基于CRISPR/Cas系统的多重基因编辑与调控技术开发及应用,针对单个细胞内实现多位点基因编辑或调控和细胞群体内实现多位点基因编辑或调控技术,依据作用原理对其进行了系统总结和阐述,包括基于CRISPR/Cas系统的双链断裂、单链断裂以及多重基因调控技术等。这些工作丰富了多重基因编辑与调控的工具,为CRISPR/Cas系统在多领域的应用作出了贡献。

The CRISPR/Cas systems comprising the clustered regularly interspaced short palindromic repeats(CRISPR)and its associated Cas protein is an acquired immune system unique to archaea or bacteria.Since its development as a gene editing tool,it has rapidly become a popular research direction in the field of synthetic biology due to its advantages of high efficiency,precision,and versatility.This technique has since revolutionized the research of many fields including life sciences,bioengineering technology,food science,and crop breeding.Currently,the single gene editing and regulation techniques based on CRISPR/Cas systems have been increasingly improved,but challenges still exist in the multiplex gene editing and regulation.This review focuses on the development and application of multiplex gene editing and regulation techniques based on the CRISPR/Cas systems,and summarizes the techniques for multiplex gene editing or regulation within a single cell or within a cell population.This includes the multiplex gene editing techniques developed based on the CRISPR/Cas systems with double-strand breaks;or with single-strand breaks;or with multiple gene regulation techniques,etc.These works have enriched the tools for the multiplex gene editing and regulation and contributed to the application of CRISPR/Cas systems in the multiple fields.