A toolbox for genetic manipulation in intestinal Clostridium symbiosum

文献类型： 外文期刊

第一作者： Yang, Pengjie

作者： Yang, Pengjie;Tian, Jinzhong;Yang, Gaohua;Gu, Yang;Jiang, Weihong;Yang, Pengjie;Tian, Jinzhong;Zhang, Lu;Ren, Yimeng;Fang, Jingyuan;Zhang, Hui;Yang, Gaohua

作者机构：

关键词： Gut Clostridium symbiosum; Toolbox; Gene overexpression; Gene deletion; Gene regulation

期刊名称：SYNTHETIC AND SYSTEMS BIOTECHNOLOGY （ 影响因子：4.8； 五年影响因子：4.9 ）

ISSN： 2405-805X

年卷期： 2024 年 9 卷 1 期

页码：

收录情况： SCI

摘要： Gut microbes are closely related with human health, but remain much to learn. Clostridium symbiosum is a conditionally pathogenic human gut bacterium and regarded as a potential biomarker for early diagnosis of intestinal tumors. However, the absence of an efficient toolbox that allows diverse genetic manipulations of this bacterium limits its in-depth studies. Here, we obtained the complete genome sequence of C. symbiosum ATCC 14940, a representative strain of C. symbiosum. On this basis, we further developed a series of genetic manipulation methods for this bacterium. Firstly, following the identification of a functional replicon pBP1 in C. symbiosum ATCC 14940, a highly efficient conjugative DNA transfer method was established, enabling the rapid introduction of exogenous plasmids into cells. Next, we constructed a dual-plasmid CRISPR/Cas12a system for genome editing in this bacterium, reaching over 60 % repression for most of the chosen genes as well as efficient deletion (>90 %) of three target genes. Finally, this toolbox was used for the identification of crucial functional genes, involving growth, synthesis of important metabolites, and virulence of C. symbiosum ATCC 14940. Our work has effectively established and optimized genome editing methods in intestinal C. symbiosum, thereby providing strong support for further basic and application research in this bacterium.

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