Zeng Sanshan, Ju Yanan, Alam Md Shah, Lu Ziwen, Hameed H M Adnan, Li Lijie, Tian Xirong, Fang Cuiting, Fang Xiange, Ding Jie, Wang Xinyue, Hu Jinxing, Wang Shuai, Zhang Tianyu
State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China.
Guangdong-Hong Kong-Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China.
mLife. 2025 Apr 23;4(2):169-180. doi: 10.1002/mlf2.70007. eCollection 2025 Apr.
, a fast-growing, non-tuberculous mycobacterium resistant to most antimicrobial drugs, causes a wide range of serious infections in humans, posing a significant public health challenge. The development of effective genetic manipulation tools for is still in progress, limiting both research and therapeutic advancements. However, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) systems have emerged as promising tools for generating highly specific double-strand breaks (DSBs) in its genome. One of the mechanisms that repair these DSBs is the error-prone nonhomologous end-joining (NHEJ) pathway, which facilitates targeted gene editing. In this study, we introduced a novel application of the CRISPR-NHEJ approach in . We demonstrated that NrgA from plays a crucial role in repairing DSBs induced by the CRISPR-Cas system in . Contrary to previous findings, our study also revealed that inhibiting or overexpressing components of homologous recombination/single-strand annealing significantly reduces the efficiency of NHEJ repair in . This discovery challenges current perspectives and suggests that NHEJ repair in may involve components from both homologous recombination and single-strand annealing pathways, highlighting the complex interactions among the three DSB repair mechanisms in .
，一种快速生长、对大多数抗菌药物耐药的非结核分枝杆菌，在人类中引起广泛的严重感染，对公共卫生构成重大挑战。针对该菌的有效基因操作工具仍在开发中，这限制了研究和治疗的进展。然而，成簇规律间隔短回文重复序列（CRISPR）相关蛋白（Cas）系统已成为在其基因组中产生高度特异性双链断裂（DSB）的有前景的工具。修复这些DSB的机制之一是易错非同源末端连接（NHEJ）途径，它有助于靶向基因编辑。在本研究中，我们介绍了CRISPR-NHEJ方法在该菌中的一种新应用。我们证明该菌的NrgA在修复由CRISPR-Cas系统诱导的DSB中起关键作用。与先前的发现相反，我们的研究还表明，抑制或过表达同源重组/单链退火的成分会显著降低该菌中NHEJ修复的效率。这一发现挑战了当前的观点，并表明该菌中的NHEJ修复可能涉及同源重组和单链退火途径的成分，突出了该菌中三种DSB修复机制之间复杂的相互作用。
