优化规律间隔成簇短回文重复序列相关核酸酶9敲入效率的研究进展

Research Progress in Optimizing Clustered Regularly Interspaced Short Palindromic Repeats-Associated Nuclease 9 Knock-in Efficiency

基因编辑技术可以通过在基因组DNA中精确插入、缺失或替换而人为地修饰靶位点的遗传物质。近年来,随着规律间隔成簇短回文重复序列及其相关核酸酶9(clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9,CRISPR/Cas9)系统的发展,研究人员对各种细胞基因组进行编辑的能力大大提高,但由于同源定向修复(homologous directed repair,HDR)效率低下,许多通过抑制非同源末端连接(non-homologous end joining,NHEJ)或增强HDR的方法,如化学调节、同步表达和同源臂优化等被开发出来用以提高基于CRISPR/Cas9系统的精确基因敲入效率,重点介绍这些方法如何优化Cas9敲入效率的最新进展。

Gene editing technology can artificially modify genetic material at target sites by inserting,deleting or replacing them precisely in genomic DNA.The development of the clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9(CRISPR/Cas9)system in recent years has greatly improved researchers,s ability to edit the genomes of various cells.However,due to the low efficiency of homologous directed repair(HDR),many approaches to improve precise gene knock-in efficiency based on the CRISPR/Cas9 system have been developed by inhibiting non-homologous end joining(NHEJ)or enhancing HDR,including chemical regulation,synchronous expression,and homologous arm optimization.Here,this review focuses on recent advances in how these methods can optimize Cas9 knock-in efficiency.