提高CRISPR/Cas9介导的动物基因组精确插入效率研究进展  

Recent developments in enhancing the efficiency of CRISPR/Cas9-mediated knock-in in animals

作　　者：李国玲 杨善欣 吴珍芳 张献伟 Guoling Li;Shanxin Yang;Zhenfang Wu;Xianwei Zhang(National Engineering Research Center for Swine Breeding Industry,College of Animal Science,South China Agricultural University,Guangzhou 510642,China;Wens Foodstuff Co.,Ltd.,Xinxing 527439,China)

机构地区：[1]华南农业大学动物科学学院,国家生猪种业工程技术研究中心,广州510642 [2]温氏食品集团股份有限公司,新兴527439

出　　处：《遗传》2020年第7期641-656,共16页Hereditas（Beijing)

基　　金：国家转基因重大专项(编号:2016ZX08006002)资助。

摘　　要：基因编辑技术是指通过人为方式在基因组插入、缺失或替换特定碱基,对遗传物质进行精确修饰和定向编辑的一种技术。近年来,锌指核酸内切酶(zinc-finger endonuclease, ZFN)、类转录激活因子效应物核酸酶(transcription activator-like effector nuclease, TALEN)、成簇规律间隔短回文重复序列及其相关系统(clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9, CRISPR/Cas9)等基因编辑技术的出现,使特异性靶向修饰动物基因组序列成为可能。虽然利用CRISPR/Cas9等基因编辑工具可以在细胞基因组高效产生双链断裂(double-strand breaks, DSB),但利用同源定向修复(homology directed repair, HDR)介导的精确插入(knock in, KI)效率却十分低下。本文结合当前基因编辑技术的发展现状,对目前提高CRISPR/Cas9介导的动物基因组KI策略进行了综述,以期为人类疾病模型制备、基因治疗和家畜遗传改良等提供借鉴。Gene-editing technology can artificially modify genetic material of targeted loci by precise insertion,deletion,or replacement in the genomic DNA.In recent years,with the developments of zinc-finger endonuclease(ZFN),transcription activator-like effector nuclease(TALEN),clustered regularly interspaced short palindromic repeats/CRISPRassociated protein 9(CRISPR/Cas9)technologies,such precise modifications of the animal genomes have become possible.Although gene-editing tools,such as CRISPR/Cas9,can efficiently generate double-strand breaks(DSBs)in mammalian cells,the homology-directed repair(HDR)mediated knock-in(KI)efficiency is extremely low.In this review,we briefly describe the current development of gene-editing tools and summarize the recent strategies to enhance the CRISPR/Cas9-mediated KI efficiency,which will provide a reference for the generation of human disease models,research on gene therapy and livestock genetic improvement.

关 键 词：基因编辑 CRISPR/Cas9 精确插入 同源定向修复 非同源末端连接 

分 类 号：Q78[生物学—分子生物学]