Genome editing of Corynebacterium glutamicum mediated with Cpf1 plus Ku/LigD

文献类型： 外文期刊

第一作者： Yang, Fa-Yu

作者： Yang, Fa-Yu;Wei, Nan;Zhang, Zhi-Hao;Wang, Mi;Liu, Ying-Chun;Zhang, Li-Fang;Gu, Feng

作者机构：

关键词： Corynebacterium glutamicum; CRISPR-Cpf1; Ku; LigD; NHEJ

期刊名称：BIOTECHNOLOGY LETTERS （ 影响因子：2.461； 五年影响因子：2.457 ）

ISSN： 0141-5492

年卷期： 2021 年 43 卷 12 期

页码：

收录情况： SCI

摘要： Objectives Corynebacterium glutamicum (C. glutamicum) has been harnessed for multi-million-ton scale production of glutamate and lysine. To further increase its amino acid production for fermentation industry, there is an acute need to develop next-generation genome manipulation tool for its metabolic engineering. All reported methods for genome editing triggered with CRISPR-Cas are based on the homologous recombination. While, it requires the generation of DNA repair template, which is a bottle-neck for its extensive application. Results In this study, we developed a method for gene knockout in C. glutamicum via CRISPR-Cpf1-coupled non-homologous end-joining (CC-NHEJ). Specifically, CRISPR-Cpf1 introduced double-strand breaks in the genome of C. glutamicum, which was further repaired by ectopically expressed two NHEJ key proteins (Mycobacterium tuberculosis Ku and ligase D). We provide the proof of concept, for CC-NHEJ, by the successful knockout of the crtYf/e gene in C. glutamicum with the efficiency of 22.00 +/- 5.56%, or something like that. Conclusion The present study reported a novel genome manipulation method for C. glutamicum.

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