CRISPR-based genome editing reveals the roles of efflux pumps in Mycobacterium abscessusOA
Mycobacterium abscessus,one of the most antimicrobial-resistant bacteria,is increasingly recognized as the cause of infections that are difficult to treat.Novel genetic manipulation tools are required to elucidate the biology,pathogenesis,and antibiotic resistance mechanisms of M.abscessus.In this study,we modified the method used to prepare M.abscessus electrocompetent cells to achieve efficient transformation,and then optimized the CRISPR-Cas9-assisted genome-editing tools to allow efficient genetic manipulation.Using these tools,we constructed 66 efflux pump mutants of M.abscessus and investigated their roles in drug resistance and virulence.We found that different efflux pumps play distinct roles in drug resistance and survival in Galleria mellonella larvae.Finally,we confirmed that MAB_2806-2807,involved in transportation of triacylglycerides,is vital for the drug resistance and virulence of M.abscessus.The molecular biology tools developed in this study will facilitate molecular research on M.abscessus.In addition,the study on efflux pumps might provide new targets for the development of new drugs and treatment regimens for M.abscessus infection.
Sishang Li;Aofei Duan;Lanyue Zhang;Chunliang Wang;Meiyi Yan;Gai-Xian Ren;Li-Ping Pan;Yi-Cheng Sun
NHC Key Laboratory of Systems Biology of Pathogens,State Key Laboratory of Respiratory Health and Multimorbidity,National Institute of Pathogen Biology and Center for Tuberculosis Research,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing,China Beijing Jishuitan Hospital,Capital Medical University,Beijing,ChinaNHC Key Laboratory of Systems Biology of Pathogens,State Key Laboratory of Respiratory Health and Multimorbidity,National Institute of Pathogen Biology and Center for Tuberculosis Research,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing,ChinaLaboratory of Molecular Biology,Beijing Key Laboratory for Drug Resistance Tuberculosis Research,Beijing Chest Hospital,Capital Medical University,Beijing Tuberculosis and Thoracic Tumor Research Institute,Beijing,ChinaNHC Key Laboratory of Systems Biology of Pathogens,State Key Laboratory of Respiratory Health and Multimorbidity,National Institute of Pathogen Biology and Center for Tuberculosis Research,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing,ChinaNHC Key Laboratory of Systems Biology of Pathogens,State Key Laboratory of Respiratory Health and Multimorbidity,National Institute of Pathogen Biology and Center for Tuberculosis Research,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing,ChinaNHC Key Laboratory of Systems Biology of Pathogens,State Key Laboratory of Respiratory Health and Multimorbidity,National Institute of Pathogen Biology and Center for Tuberculosis Research,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing,ChinaLaboratory of Molecular Biology,Beijing Key Laboratory for Drug Resistance Tuberculosis Research,Beijing Chest Hospital,Capital Medical University,Beijing Tuberculosis and Thoracic Tumor Research Institute,Beijing,ChinaNHC Key Laboratory of Systems Biology of Pathogens,State Key Laboratory of Respiratory Health and Multimorbidity,National Institute of Pathogen Biology and Center for Tuberculosis Research,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing,China
医药卫生
CRISPR-Cas9drug resistanceefflux pumpgenome editingMycobacterium abscessus
《mLife》 2026 (1)
P.68-82,15
supported by the National Key R&D Program of China(2022YFC2303200)the National Natural Science Foundation of China(32370199)the CAMS Initiative for Innovative Medicine(2021-I2M-1-035)the Beijing Jishuitan Research Funding(ZR-202520).
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