CRISPR/Cas9基因组编辑技术及其在链霉菌中的应用

doi:10.13560/j.cnki.biotech.bull.1985.2017-0877

摘要/Abstract

摘要： 链霉菌（Streptomyces）以产生多种抗生素而著称,长期以来都是生命科学与医学、药学及工业领域的研究热点。然而,链霉菌的遗传操作十分困难,传统的基因编辑手段因效率低、实验周期长及操作步骤繁琐等原因正逐渐被素有“魔剪”之称的CRISPR（Clustered regularly interspaced short palindromic repeats）/Cas9（CRISPR-associated protein 9）系统所替代。该技术可以对基因组特定位点进行靶向编辑,包括缺失（Indel）、修复（Repair）和替换（Replacement）等。通过比较目前在链霉菌中基因编辑手段,总结出CRISPR/Cas9系统在链霉菌应用方面具有操作简单、效率高、成本低、实验周期短,以及能够同时敲除多个基因等优势,并综述了该系统在链霉菌中的应用,以及汇总了CRISPR/Cas9工具箱的靶向空间,最后针对该系统在链霉菌定向育种方面及合成新的抗生素方面进行展望,以期为相关领域的工作提供参考。

关键词: CRISPR/Cas9, 基因组编辑, 特异性, 高效性, 链霉菌

Abstract: Streptomyces are known to produce a variety of antibiotics and have long been a research focus in life sciences and medicine,pharmacy,and industry. However,the genetic manipulation of Streptomyces is very difficult,thus the traditional means of genetic editing due to low efficiency,long experimental cycle and cumbersome steps and other reasons are gradually replaced by CRISPR/Cas9 system called as “magic scissors”. This technique allows for targeted editing of genome-specific sites,including indel,repair and replacement. By comparing the current gene editing methods in Streptomyces,it is concluded that CRISPR/Cas9 system has the advantages of simple operation,high efficiency,low cost,short experimental cycle and the ability to simultaneously knock out multiple genes in the application of Streptomyces. We also review the application of the system in Streptomyces and summarize the target space of CRISPR/Cas9 toolbox. Finally,we prospect the potential of CRISPR/Cas9 system in Streptomyces orientated-breeding and synthetizing new antibiotics,aiming at providing references for the work of related fields.

Key words: CRISPR/Cas9, genome editing, specificity, high efficiency, Streptomyces

乔龙亮,庞建虎,党晨阳,黄海龙,朱鹏. CRISPR/Cas9基因组编辑技术及其在链霉菌中的应用[J]. 生物技术通报, 2018, 34(5): 32-40.

QIAO Long-liang, PANG Jian-hu, DANG Chen-yang, HUANG Hai-long, ZHU Peng. CRISPR/Cas9 Genome Editing Technology and Its Application in Streptomyces[J]. Biotechnology Bulletin, 2018, 34(5): 32-40.

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