Tuberculosis(TB)is a formidable global health problem and ranks above HIV as the leading cause of death world wide.In2017,a total number of 10.0 million cases of TB were reported, which resulted in1.3 million TB death...Tuberculosis(TB)is a formidable global health problem and ranks above HIV as the leading cause of death world wide.In2017,a total number of 10.0 million cases of TB were reported, which resulted in1.3 million TB deaths. Resistance to standard anti-TB展开更多
Treatment of Mycobacterium abscessus(Mab)infections is very challenging due to its intrinsic resistance to most available drugs.Therefore,it is crucial to discover novel anti-Mab drugs.In this study,we explored an int...Treatment of Mycobacterium abscessus(Mab)infections is very challenging due to its intrinsic resistance to most available drugs.Therefore,it is crucial to discover novel anti-Mab drugs.In this study,we explored an intrinsic resistance mechanism through which Mab resists echinomycin(ECH).ECH showed activity against Mab at a minimum inhibitory concentration(MIC)of 2μg/ml.A embC strain in which the embC gene was knocked out showed hypersensitivity to ECH(MIC:0.0078-0.0156μg/ml).The MICs of ECH-resistant strains screened with reference to AembC ranged from 0.25 to 1μg/ml.Mutations in EmbB,including D306A,D306N,R350G,V555l,and G581S,increased the Mab's resistance to ECH when overexpressed in AembC individually(MIC:0.25-0.5μg/ml).These EmbB mutants,edited using the CRISPR/Cpf1 system,showed heightened resistance to ECH(MIC:0.25-0.5μg/ml).The permeability of these Mab strains with edited genes and overexpression was reduced,as evidenced by an ethidium bromide accumulation assay,but it remained significantly higher than that of the parent Mab.In summary,our study demonstrates that ECH exerts potent anti-Mab activity and confirms that EmbB and EmbC are implicated in Mab's sensitivity to ECH.Mutation in EmbB may partially compensate foralossof EmbCfunction.展开更多
Mycobacterium tuberculosis, a Gram-positive bacterium of great clinical relevance, is a lethal pathogen owing to its complex physiological characteristics and development of drug resistance. Several molecular genetic ...Mycobacterium tuberculosis, a Gram-positive bacterium of great clinical relevance, is a lethal pathogen owing to its complex physiological characteristics and development of drug resistance. Several molecular genetic tools have been developed in the past few decades to study this microorganism. These tools have been instrumental in understanding how M. tuberculosis became a successful pathogen. Advanced molecular genetic tools have played a significant role in exploring the complex pathways involved in M. tuberculosis pathogenesis. Here, we review various molecular genetic tools used in the study of M. tuberculosis. Further, we discuss the applications of clustered regularly interspaced short palindromic repeat interference（CRISPRi）, a novel technology recently applied in M. tuberculosis research to study target gene functions. Finally, prospective outcomes of the applications of molecular techniques in the field of M. tuberculosis genetic research are also discussed.展开更多
基金supported by Guangzhou Municipal Industry and Research Collaborative Innovation Program [201508020248,201604020019]Guangzhou Municipal Clinical Medical Center Program [155700012]+3 种基金partially by the Key Project Grant [SKLRD2016ZJ003] from the State Key Lab of Respiratory Disease,Guangzhou Institute of Respiratory DiseasesScience and Technology Innovation Leader of Guangdong Province [2016TX03R095,to TZ]by the UCAS Fellowship Program(to H.M.A.H)by the CAS-TWAS President’s Fellowship(to G.C)
文摘Tuberculosis(TB)is a formidable global health problem and ranks above HIV as the leading cause of death world wide.In2017,a total number of 10.0 million cases of TB were reported, which resulted in1.3 million TB deaths. Resistance to standard anti-TB
基金This work was supported by the National Key R&D Program of China(2021YFA1300900)the National Natural Science Foundation of China(21920102003,82022067,and 22037006)+5 种基金the Chinese Academy of Sciences Grants(154144KYSB20190005 and YJKYYQ20210026)the Key R&D Program of Sichuan Provenience(2023YFSY0047)the State Key Laboratory of Respiratory Disease,Guangzhou Institute of Respiratory Diseases,First Affiliated Hospital of Guangzhou Medical University(SKLRD-Z-202414,SKLRD-OP-202324,SKLRD-Z-202301,SKLRD-OP-202113,and SKLRD-Z-202412)Guangzhou Scienceaand Technology Plan-Youth Doctoral"Sail"Project(2024A04J4273)President's International Fellowship Initiative-CAS(2023VBC0015)the National Foreign Young Talent Program(QN2022031002L).
文摘Treatment of Mycobacterium abscessus(Mab)infections is very challenging due to its intrinsic resistance to most available drugs.Therefore,it is crucial to discover novel anti-Mab drugs.In this study,we explored an intrinsic resistance mechanism through which Mab resists echinomycin(ECH).ECH showed activity against Mab at a minimum inhibitory concentration(MIC)of 2μg/ml.A embC strain in which the embC gene was knocked out showed hypersensitivity to ECH(MIC:0.0078-0.0156μg/ml).The MICs of ECH-resistant strains screened with reference to AembC ranged from 0.25 to 1μg/ml.Mutations in EmbB,including D306A,D306N,R350G,V555l,and G581S,increased the Mab's resistance to ECH when overexpressed in AembC individually(MIC:0.25-0.5μg/ml).These EmbB mutants,edited using the CRISPR/Cpf1 system,showed heightened resistance to ECH(MIC:0.25-0.5μg/ml).The permeability of these Mab strains with edited genes and overexpression was reduced,as evidenced by an ethidium bromide accumulation assay,but it remained significantly higher than that of the parent Mab.In summary,our study demonstrates that ECH exerts potent anti-Mab activity and confirms that EmbB and EmbC are implicated in Mab's sensitivity to ECH.Mutation in EmbB may partially compensate foralossof EmbCfunction.
基金supported by the National Mega-project of China for Innovative Drugs(2018ZX09721001-003-003)for Main Infectious Diseases(2017ZX10302301-003-002)+7 种基金the National Natural Science Foundation of China(No.81572037)the grants of Chinese Academy of Sciences(154144KYSB20150045,YJKYYQ20170036,KFZD-SW-207)the Public Research and Capacity Building Project of Guangdong Province(2017A020212004)partially supported by Guangzhou Municipal Industry and Research Collaborative Innovation Program(201508020248,201604020019)the Key Project(SKLRD2016ZJ003)from the State Key Lab of Respiratory Disease,Guangzhou Institute of Respiratory Diseases,First Affiliated Hospital of Guangzhou Medical UniversityT.Z.received support“Science and Technology Innovation Leader of Guangdong Province(2016TX03R095)”CAS-TWAS President's Ph D Fellowship Program(to C.C.and M.M.I.)UCAS Ph D Fellowship Program(to H.M.A.H.and J.M.)for International Students
文摘Mycobacterium tuberculosis, a Gram-positive bacterium of great clinical relevance, is a lethal pathogen owing to its complex physiological characteristics and development of drug resistance. Several molecular genetic tools have been developed in the past few decades to study this microorganism. These tools have been instrumental in understanding how M. tuberculosis became a successful pathogen. Advanced molecular genetic tools have played a significant role in exploring the complex pathways involved in M. tuberculosis pathogenesis. Here, we review various molecular genetic tools used in the study of M. tuberculosis. Further, we discuss the applications of clustered regularly interspaced short palindromic repeat interference（CRISPRi）, a novel technology recently applied in M. tuberculosis research to study target gene functions. Finally, prospective outcomes of the applications of molecular techniques in the field of M. tuberculosis genetic research are also discussed.
