{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T01:29:13Z","timestamp":1787016553645,"version":"build-2736575974"},"reference-count":50,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T00:00:00Z","timestamp":1784764800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T00:00:00Z","timestamp":1784764800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["GM103429"],"award-info":[{"award-number":["GM103429"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000072","name":"National Institute of Dental and Craniofacial Research","doi-asserted-by":"publisher","award":["DE033403"],"award-info":[{"award-number":["DE033403"]}],"id":[{"id":"10.13039\/100000072","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000072","name":"National Institute of Dental and Craniofacial Research","doi-asserted-by":"publisher","award":["DE034550"],"award-info":[{"award-number":["DE034550"]}],"id":[{"id":"10.13039\/100000072","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2026,7,23]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Streptococcus mutans<\/jats:italic>\n                      is a primary architect of dental caries, utilizing complex genetic networks to build resilient, acid-producing biofilms. While pooled screens (Tn-seq) have identified important fitness factors, they often fail to capture extracellular or moderate-effect determinants due to community-level masking. Therefore, to study biofilm phenotypes, we constructed a comprehensive arrayed library of 9,216 mutants and used Cartesian Pooling-Coordinate Sequencing (CP-CSeq) to establish a sequence-defined resource covering 51% of non-essential genes. By screening the entire collection in isolation, we identified several novel biofilm determinants, including the putative metal transporter SMU_635 and the glycosylation-associated protein SMU_2160. However, systematic whole-genome sequencing (WGS) of our hits revealed an interesting level of genomic instability: 25% of biofilm-defective mutants had undergone spontaneous recombination at the\n                      <jats:italic toggle=\"yes\">gtfBC<\/jats:italic>\n                      locus, while 7% had lost the Tn\n                      <jats:italic toggle=\"yes\">Smu1<\/jats:italic>\n                      element, an excision rate 1,000-fold higher than previously reported. While targeted mutagenesis confirmed that Tn\n                      <jats:italic toggle=\"yes\">Smu1<\/jats:italic>\n                      loss does not impact biofilm integrity, the\n                      <jats:italic toggle=\"yes\">gtfBC<\/jats:italic>\n                      deletions directly accounted for the most severe phenotypes, highlighting a systemic risk of misattributing gene functions to primary transposon insertions. Our findings provide a powerful new genetic resource for the\n                      <jats:italic toggle=\"yes\">S. mutans<\/jats:italic>\n                      community while establishing a critical new standard: an arrayed library is only as defined as its underlying genome, making systematic genomic verification an essential prerequisite for accurate functional genomics.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Streptococcus mutans<\/jats:italic>\n                      causes dental caries through resilient, acidogenic biofilm formation. While pooled screens often overlook extracellular or moderate-effect determinants due to community masking, this study presents a sequence-defined arrayed mutant library to dissect individual gene functions in isolation. Beyond known machinery, we identified novel biofilm determinants, including metal transporter SMU_635 and glycosylation-associated protein SMU_2160. Crucially, we uncovered pervasive genomic instability at the\n                      <jats:italic toggle=\"yes\">gtfBC<\/jats:italic>\n                      and Tn\n                      <jats:italic toggle=\"yes\">Smu1<\/jats:italic>\n                      loci. This reveals a systemic functional genomics risk: misattributing phenotypes to primary mutations when backgrounds undergo large-scale rearrangements. By establishing whole-genome verification as a necessary standard, this research ensures that future therapeutic target identification is built upon a verified genetic foundation.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/jb.00174-26","type":"journal-article","created":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T12:00:31Z","timestamp":1782129631000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Genomic instability and biofilm determinants in\n                    <i>Streptococcus mutans<\/i>\n                    : insights from a sequence-defined arrayed transposon library"],"prefix":"10.1128","volume":"208","author":[{"given":"Ana Karen","family":"Solano Morales","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/006pyvd89","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biological Sciences, Beck College of Sciences and 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