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有机化学
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有机化学 ›› 2021, Vol. 41 ›› Issue (11): 4279-4288.DOI: 10.6023/cjoc202105035 上一篇    下一篇

综述与进展

CRISPR/Cas9基因编辑系统在获取放线菌天然产物中的应用

乔怡a, 张庆林b,c, 陈单丹b,c,*(), 刘美娜a,*(), 刘文b,c,*()   

  1. a 上海应用技术大学化学与环境工程学院 上海 201418
    b 中国科学院上海有机化学研究所 生命有机化学国家重点实验室 上海 200032
    c 中国科学院上海有机化学研究所 湖州生物制造创新中心 浙江湖州 313000
  • 收稿日期:2021-05-20 修回日期:2021-06-16 发布日期:2021-07-05
  • 通讯作者: 陈单丹, 刘美娜, 刘文
  • 基金资助:
    国家自然科学基金(21977109); 国家自然科学基金(32030002); 国家自然科学基金(21750004); 国家重点研发计划(2019YFA0905400); 中国科学院战略性先导科技专项(XDB20020200); 中国科学院青年创新促进会(2017303)

Application of CRISPR/Cas9 Gene Editing System in Obtaining Natural Products in Actinomycetes

Yi Qiaoa, Qinglin Zhangb,c, Dandan Chenb,c(), Meina Liua(), Wen Liub,c()   

  1. a School of Chemical and Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418
    b State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032
    c Huzhou Center of Bio-synthetic Innovation, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Huzhou, Zhejiang 313000
  • Received:2021-05-20 Revised:2021-06-16 Published:2021-07-05
  • Contact: Dandan Chen, Meina Liu, Wen Liu
  • Supported by:
    National Natural Science Foundation of China(21977109); National Natural Science Foundation of China(32030002); National Natural Science Foundation of China(21750004); National Key R&D Program of China(2019YFA0905400); Strategic Priority Research Program of the Chinese Academy of Sciences(XDB20020200); Youth Innovation Promotion Association of the Chinese Academy of Sciences(2017303)

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作为临床药物的重要来源, 放线菌天然产物及其衍生物已被大规模开发并应用于临床. 为了实现放线菌来源活性天然产物的高效合成、新型天然产物的结构衍生以及未知天然产物的定向挖掘, 往往需要聚焦于生物合成基因簇, 运用生物学技术开展理性的遗传改造. 规律性成簇间隔的短回文重复序列及相关蛋白(CRISPR/Cas)是部分细菌和古细菌在长期自然进化过程中发展而来的一种对抗外源遗传物质入侵的适应性免疫防御系统. 以CRISPR/Cas9为代表的基因编辑系统已被广泛应用于脱氧核糖核酸(DNA)片段的靶向特异性切割, 为生物技术领域的发展提供了无限可能. 综述了CRISPR/Cas9系统在放线菌遗传改造中的应用情况, 包括在基因簇抓取、编辑与重构、原位同源重组以及代谢调控方面的研究进展, 涉及全新的CRISPR/Cas9介导的基因编辑方法以及代表性的成功案例, 为高效利用CRISPR/Cas9系统获取放线菌天然产物提供了借鉴.

关键词: 放线菌, 天然产物, 生物合成基因簇, 基因编辑, 规律性间成簇间隔的短回文重复序列及相关蛋白9 (CRISPR/Cas9)

As important source of clinical drugs, the natural products in actinomycetes and their derivatives have been developed on a large scale and used in clinical practice. In order to achieve efficient synthesis of active natural products, structural derivation of new natural products, and targeted mining of unknown natural products, researches mainly focus on the biosynthetic gene clusters from actinomycetes, which should be rationally engineered by advanced biotechnology. CRISPR/ Cas, which represents the clustered regularly interspaced short palindromic repeats and the associated protein, is an adaptive immune system developed by some bacteria and archaea in the long-term natural evolution process to resist the invasion of foreign genetic materials. As a representative of gene editing system, the clustered regularly interspaced short palindromic repeats and the associated protein 9 (CRISPR/Cas9) has been widely used in the targeted editing of deoxyribonucleic acid (DNA) fragments, which provides unlimited possibilities for the development of biotechnology. This paper reviews the application of the CRISPR/Cas9 system in the genetic manipulation of actinomycetes, including the recent research progress in capture, editing and reconstruction of gene clusters, in situ homologous recombination and metabolic regulation, which involves the novel CRISPR/Cas9-mediated gene editing methods and the representative successful cases. This paper will provide valuable information for the efficient use of CRISPR/Cas9 to obtain natural products in actinomycetes.

Key words: actinomycetes, natural products, biosynthetic gene clusters, gene editing, the clustered regularly interspaced short palindromic repeats and the associated protein 9 (CRISPR/Cas9)