{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T09:04:25Z","timestamp":1790845465415,"version":"4.1.0"},"reference-count":44,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2024,6,4]],"date-time":"2024-06-04T00:00:00Z","timestamp":1717459200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,6,4]],"date-time":"2024-06-04T00:00:00Z","timestamp":1717459200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100008076","name":"Ohio University","doi-asserted-by":"publisher","award":["START UP, RSAC"],"award-info":[{"award-number":["START UP, RSAC"]}],"id":[{"id":"10.13039\/100008076","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2024,6,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              <jats:italic>Clostridioides difficile<\/jats:italic>\n              infection (CDI) with high morbidity and high mortality is an urgent threat to public health, and\n              <jats:italic>C. difficile<\/jats:italic>\n              pathogenesis studies are eagerly required for CDI therapy. The major surface layer protein, SlpA, was supposed to play a key role in\n              <jats:italic>C. difficile<\/jats:italic>\n              pathogenesis; however, a lack of isogenic\n              <jats:italic>slpA<\/jats:italic>\n              mutants has greatly hampered analysis of SlpA functions. In this study, the whole\n              <jats:italic>slpA<\/jats:italic>\n              gene was successfully deleted for the first time via CRISPR-Cas9 system. Deletion of\n              <jats:italic>slpA<\/jats:italic>\n              in\n              <jats:italic>C. difficile<\/jats:italic>\n              resulted in smaller, smother-edged colonies, shorter bacterial cell size, and aggregation in suspension. For life cycle, the mutant demonstrated lower growth (changes of optical density at 600 nm, OD600) but higher cell density (colony-forming unit, CFU), decreased toxins production, and inhibited sporulation. Moreover, the mutant was more impaired in motility, more sensitive to vancomycin and Triton X-100-induced autolysis, releasing more lactate dehydrogenase. In addition, SlpA deficiency led to robust biofilm formation but weak adhesion to human host cells.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                <jats:italic>Clostridioides difficile<\/jats:italic>\n                infection (CDI) has been the most common hospital-acquired infection, with a high rate of antibiotic resistance and recurrence incidences, become a debilitating public health threat. It is urgently needed to study\n                <jats:italic>C. difficile<\/jats:italic>\n                pathogenesis for developing efficient strategies as CDI therapy. SlpA was indicated to play a key role in\n                <jats:italic>C. difficile<\/jats:italic>\n                pathogenesis. However, analysis of SlpA functions was hampered due to lack of isogenic\n                <jats:italic>slpA<\/jats:italic>\n                mutants. Surprisingly, the first\n                <jats:italic>slpA<\/jats:italic>\n                deletion\n                <jats:italic>C. difficile<\/jats:italic>\n                strain was generated in this study via CRISPR-Cas9, further negating the previous thought about\n                <jats:italic>slpA<\/jats:italic>\n                being essential. Results in this study will provide direct proof for roles of SlpA in\n                <jats:italic>C. difficile<\/jats:italic>\n                pathogenesis, which will facilitate future investigations for new targets as vaccines, new therapeutic agents, and intervention strategies in combating CDI.\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/spectrum.04005-23","type":"journal-article","created":{"date-parts":[[2024,5,6]],"date-time":"2024-05-06T13:00:40Z","timestamp":1715000440000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":17,"title":["Revealing roles of S-layer protein (SlpA) in\n            <i>Clostridioides difficile<\/i>\n            pathogenicity by generating the first\n            <i>slpA<\/i>\n            gene deletion mutant"],"prefix":"10.1128","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8548-1602","authenticated-orcid":true,"given":"Shaohua","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Biomedical Sciences, Ohio University Heritage College of Osteopathic Medicine, Ohio University, Athens, Ohio, USA"},{"name":"Infectious and Tropical Disease Institute, Ohio University, Athens, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria C.","family":"Courreges","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, Ohio University Heritage College of Osteopathic Medicine, Ohio University, Athens, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingjun","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering, Institute for Corrosion and Multiphase Technology, Ohio University, Athens, Ohio, 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