遗传

• 综述 •    

提高CRISPR/Cas9介导的动物基因组精确插入效率研究进展

李国玲1,杨善欣1,吴珍芳2,张献伟3   

  1. 1. 华南农业大学
    2. 华南农业大学动物科学院
    3. 广东温氏食品集团股份有限公司
  • 收稿日期:2020-03-04 修回日期:2020-04-24 出版日期:2020-06-01 发布日期:2020-06-01
  • 通讯作者: 张献伟
  • 基金资助:
    国家转基因重大专项

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

  • Received:2020-03-04 Revised:2020-04-24 Online:2020-06-01 Published:2020-06-01

摘要: 基因编辑技术是指人为方式在基因组插入、缺失或替换特定碱基对等,对遗传物质进行精确修饰和定向编辑的一种技术。近年来,随着锌指核酸内切酶(zinc-finger endonuclease, ZFN)、类转录激活因子效应物核酸酶(transcription activator-like effector nuclease, TALEN)、规律成簇的间隔短回文重复(clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins 9, CRISPR/Cas9)等基因编辑技术的出现,使得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。

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

Abstract: Gene-editing technology can artificially modify genetic material and perform targeted loci by precise insertion, deletion, or replacement in genomic DNA. In recent years, with the development of zinc-finger endonuclease (ZFN), transcription activator-like effector nuclease (TALEN), clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins 9 (CRISPR/Cas9), the precise modification has 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 awfully low. In this review, we will briefly describe the current development of gene-editing tools. And the recent strategies in order to enhance the CRISPR/Cas9-mediated KI efficiency are furtherly summarized, which will provide a reference for the generation of human disease models, gene therapy and livestock genetic improvement.

Key words: gene editing, CRISPR / Cas9, knock in, homology directed repair, non-homologous end joining