{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T18:34:11Z","timestamp":1790102051059,"version":"4.0.1"},"reference-count":40,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2025,4,23]],"date-time":"2025-04-23T00:00:00Z","timestamp":1745366400000},"content-version":"vor","delay-in-days":22,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["mLife"],"published-print":{"date-parts":[[2025,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Mycobacterium abscessus<\/jats:italic>\n                    , a fast\u2010growing, non\u2010tuberculous 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\n                    <jats:italic>M. abscessus<\/jats:italic>\n                    is still in progress, limiting both research and therapeutic advancements. However, the clustered regularly interspaced short palindromic repeats (CRISPR)\u2010associated protein (Cas) systems have emerged as promising tools for generating highly specific double\u2010strand breaks (DSBs) in its genome. One of the mechanisms that repair these DSBs is the error\u2010prone nonhomologous end\u2010joining (NHEJ) pathway, which facilitates targeted gene editing. In this study, we introduced a novel application of the CRISPR\u2010NHEJ approach in\n                    <jats:italic>M. abscessus<\/jats:italic>\n                    . We demonstrated that NrgA from\n                    <jats:italic>M. marinum<\/jats:italic>\n                    plays a crucial role in repairing DSBs induced by the CRISPR\u2010Cas system in\n                    <jats:italic>M. abscessus<\/jats:italic>\n                    . Contrary to previous findings, our study also revealed that inhibiting or overexpressing components of homologous recombination\/single\u2010strand annealing significantly reduces the efficiency of NHEJ repair in\n                    <jats:italic>M. abscessus<\/jats:italic>\n                    . This discovery challenges current perspectives and suggests that NHEJ repair in\n                    <jats:italic>M. abscessus<\/jats:italic>\n                    may involve components from both homologous recombination and single\u2010strand annealing pathways, highlighting the complex interactions among the three DSB repair mechanisms in\n                    <jats:italic>M. abscessus<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1002\/mlf2.70007","type":"journal-article","created":{"date-parts":[[2025,4,23]],"date-time":"2025-04-23T03:10:16Z","timestamp":1745377816000},"page":"169-180","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A CRISPR\u2010nonhomologous end\u2010joining\u2010based strategy for rapid and efficient gene disruption in\n                    <i>Mycobacterium abscessus<\/i>"],"prefix":"10.1002","volume":"4","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-2047-7669","authenticated-orcid":false,"given":"Sanshan","family":"Zeng","sequence":"first","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"University of Chinese Academy of Sciences Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanan","family":"Ju","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"School of Basic Medical Sciences, Division of Life Science and Medicine University of Science and Technology of China Hefei China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Shah","family":"Alam","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"University of Chinese Academy of Sciences Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziwen","family":"Lu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"University of Chinese Academy of Sciences Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1993-9547","authenticated-orcid":false,"given":"H. M. Adnan","family":"Hameed","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lijie","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"School of Basic Medical Sciences, Division of Life Science and Medicine University of Science and Technology of China Hefei China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xirong","family":"Tian","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"University of Chinese Academy of Sciences Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cuiting","family":"Fang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"University of Chinese Academy of Sciences Beijing China"},{"name":"State Key Laboratory of Respiratory Disease Guangzhou Chest Hospital Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiange","family":"Fang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"University of Chinese Academy of Sciences Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Ding","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Institute of Physical Science and Information Technology Anhui University Hefei China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyue","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease Guangzhou Chest Hospital Guangzhou China"},{"name":"Guangzhou National Laboratory Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinxing","family":"Hu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease Guangzhou Chest Hospital Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuai","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"State Key Laboratory of Respiratory Disease Guangzhou Chest Hospital Guangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tianyu","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Respiratory Disease, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"Guangdong\u2010Hong Kong\u2010Macao Joint Laboratory of Respiratory Infectious Diseases, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"China\u2010New Zealand Joint Laboratory on Biomedicine and Health, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China"},{"name":"University of Chinese Academy of Sciences Beijing China"},{"name":"School of Basic Medical Sciences, Division of Life Science and Medicine University of Science and Technology of China Hefei China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,4,23]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.659997"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1186\/s40249-021-00844-1"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1136\/thoraxjnl-2015-207983"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1164\/rccm.200604-571ST"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkr578"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2016.06.005"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.09.038"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2016.184"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tcb.2015.07.009"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.MGM2-0024-2013"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tig.2021.02.008"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.000578"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2010.07463.x"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1042\/BST0370539"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro1768"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth996"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00947-17"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02364-19"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.02809-23"},{"key":"e_1_2_10_21_1","doi-asserted-by":"crossref","DOI":"10.1128\/spectrum.02763-21","article-title":"Arabinosyltransferase C mediates multiple drugs intrinsic resistance by altering cell envelope permeability in Mycobacterium abscessus","volume":"10","author":"Wang S","year":"2022","journal-title":"Microbiol Spectr"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2021.633510"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2021.09.051"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-47312-3"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2020.140468"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01179-14"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.304741"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M103523200"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi0340548"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2004.04102.x"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks1298"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M114.585117"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciz071"},{"key":"e_1_2_10_34_1","first-page":"Unit 10D.2","article-title":"Genetic manipulation of Mycobacterium abscessus","volume":"10","author":"Medjahed H","year":"2010","journal-title":"Curr Protoc Microbiol"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2018.00069"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-023-02214-y"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw223"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1631908"},{"key":"e_1_2_10_39_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jproteome.6b00116"},{"key":"e_1_2_10_40_1","doi-asserted-by":"publisher","DOI":"10.1534\/g3.119.400737"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-2450-9_7"}],"container-title":["mLife"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/mlf2.70007","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,8,11]],"date-time":"2025-08-11T09:40:00Z","timestamp":1754905200000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/mlf2.70007"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4]]},"references-count":40,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2025,4]]}},"alternative-id":["10.1002\/mlf2.70007"],"URL":"https:\/\/doi.org\/10.1002\/mlf2.70007","archive":["Portico"],"relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2024.01.29.577284","asserted-by":"object"}]},"ISSN":["2097-1699","2770-100X"],"issn-type":[{"value":"2097-1699","type":"print"},{"value":"2770-100X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4]]},"assertion":[{"value":"2024-08-28","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-01-13","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-04-23","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}