遗传 ›› 2020, Vol. 42 ›› Issue (7): 641-656.doi: 10.16288/j.yczz.20-056

• 综述 • 上一篇    下一篇

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1,2, 张献伟2()   

  1. 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
  • 收稿日期:2020-03-04 修回日期:2020-04-24 出版日期:2020-07-20 发布日期:2020-06-01
  • 通讯作者: 张献伟 E-mail:zxianw@163.com
  • 作者简介:李国玲,在读博士研究生,专业方向:基因编辑。E-mail: 792268184@qq.com
  • 基金资助:
    国家转基因重大专项资助编号(2016ZX08006002)

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

Guoling Li1, Shanxin Yang1, Zhenfang Wu1,2, Xianwei Zhang2()   

  1. 1. National Engineering Research Center for Swine Breeding Industry, College of Animal Science, South China Agricultural University, Guangzhou 510642, China
    2. Wens Foodstuff Co., Ltd., Xinxing 527439, China
  • Received:2020-03-04 Revised:2020-04-24 Online:2020-07-20 Published:2020-06-01
  • Contact: Zhang Xianwei E-mail:zxianw@163.com
  • Supported by:
    Supported by the National Transgenic Major Projects No(2016ZX08006002)

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。

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 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/CRISPR- associated 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.

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