{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T10:28:21Z","timestamp":1784802501631,"version":"3.55.0"},"reference-count":25,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2025,9,23]],"date-time":"2025-09-23T00:00:00Z","timestamp":1758585600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009238","name":"Virginia Commonwealth University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100009238","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R21AI166168"],"award-info":[{"award-number":["R21AI166168"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R25GM089614"],"award-info":[{"award-number":["R25GM089614"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2025,12]]},"abstract":"ABSTRACT<\/jats:title>\n \n Sneathia vaginalis<\/jats:italic>\n is a common component of the vaginal microbiome and is emerging as a marker for preterm birth. It produces the cytopathogenic toxin A (CptA), which is capable of lysing human red blood cells and permeabilizing epithelial cells. However, the role of CptA and other potential virulence factors in pathogenesis has been difficult to characterize due to the lack of genetic tools for targeted deletion in\n \n S.\u2009vaginalis<\/jats:italic>\n <\/jats:styled-content>\n . The objective of this study was to create the first isogenic gene deletion mutant in\n \n S.\u2009vaginalis<\/jats:italic>\n <\/jats:styled-content>\n . We chose the\n cptA<\/jats:italic>\n gene as a target for deletion because of its role in virulence. We characterized the restriction\u2010modification profile in\n \n S.\u2009vaginalis<\/jats:italic>\n <\/jats:styled-content>\n to increase the chances that exogenous DNA would resist restriction digestion, and we identified an antibiotic resistance cassette that is functional in this species. We identified a genetic locus encoding a Dam methylase and a restriction endonuclease with DpnII\u2010like activity in\n \n S.\u2009vaginalis<\/jats:italic>\n <\/jats:styled-content>\n strain SN35. By convention, this newly described restriction endonuclease would be named SvaSI for\n \n S.\u2009vaginalis<\/jats:italic>\n <\/jats:styled-content>\n SN35. Using plasmid DNA purified from a Dam+\n \n E.\u2009coli<\/jats:italic>\n <\/jats:styled-content>\n strain to evade SvaSI restriction, we successfully replaced\n cptA<\/jats:italic>\n with an erythromycin resistance cassette encoding the\n ermF<\/jats:italic>\n and\n ermAM<\/jats:italic>\n genes, creating the first genetically engineered deletion mutation in this species. Results revealed that CptA is necessary for the hemolytic and cytopathogenic activities of\n \n S.\u2009vaginalis<\/jats:italic>\n <\/jats:styled-content>\n . This work is a resource that lays the foundation for the development of additional genetic tools for\n \n S.\u2009vaginalis<\/jats:italic>\n <\/jats:styled-content>\n and facilitates the characterization of additional genes in this emerging pathogen.\n <\/jats:p>","DOI":"10.1111\/mmi.70024","type":"journal-article","created":{"date-parts":[[2025,9,23]],"date-time":"2025-09-23T10:29:12Z","timestamp":1758623352000},"page":"481-490","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Targeted Deletion of the Cytopathogenic Toxin A Gene in\n Sneathia vaginalis<\/i>"],"prefix":"10.1111","volume":"124","author":[{"given":"Rishi M.","family":"Ray","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology Virginia Commonwealth University Richmond Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Phoebe V.","family":"Bridy","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology Virginia Commonwealth University Richmond Virginia USA"},{"name":"Philips Institute for Oral Health Research, School of Dentistry Richmond Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aubree G.","family":"Musicant","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology Virginia Commonwealth University Richmond Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiny","family":"Chandravel","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology Virginia Commonwealth University Richmond Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youstina Y.","family":"Aziz","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology Virginia Commonwealth University Richmond Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jasmine C.","family":"Cruz","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology Virginia Commonwealth University Richmond Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1052-1988","authenticated-orcid":false,"given":"Kimberly K.","family":"Jefferson","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology Virginia Commonwealth University Richmond Virginia USA"},{"name":"Center for Microbiome Engineering and Data Analysis Virginia Commonwealth University Richmond Virginia USA"},{"name":"Department of Obstetrics and Gynecology Virginia Commonwealth University Richmond Virginia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,9,23]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-009-2349-1"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.siny.2011.10.001"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1515\/jpm.2010.078"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0003056"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1099\/ijsem.0.004663"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-019-0450-2"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.63.4.1521-1528.1995"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1055\/s-2004-831884"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00162-20"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01206-08"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2164-13-S8-S4"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkr1321"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00265063"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1820256116"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.plasmid.2005.06.002"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.00392"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1462-5822.2001.00123.x"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(89)90300-0"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkg274"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.3109\/14767058.2014.958463"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkx965"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12275-021-0674-y"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.40.9.3346-3349.2002"},{"key":"e_1_2_8_25_1","doi-asserted-by":"crossref","unstructured":"Sun H. A. P.Luu M.Danielson T. T.Vo andT. P.Burke.2023.\u201cHost Glutathione Is Required forRickettsia parkerito Properly Septate Avoid Ubiquitylation and Survive in Macrophages.biorxiv.\u201dhttps:\/\/doi.org\/10.1101\/2023.10.02.560592v1.","DOI":"10.1101\/2023.10.02.560592"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkz841"}],"container-title":["Molecular Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/mmi.70024","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T06:07:25Z","timestamp":1764828445000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/mmi.70024"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,23]]},"references-count":25,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2025,12]]}},"alternative-id":["10.1111\/mmi.70024"],"URL":"https:\/\/doi.org\/10.1111\/mmi.70024","archive":["Portico"],"relation":{},"ISSN":["0950-382X","1365-2958"],"issn-type":[{"value":"0950-382X","type":"print"},{"value":"1365-2958","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,23]]},"assertion":[{"value":"2025-05-22","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-09-11","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-09-23","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}