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Inspection of all\n                      <jats:italic toggle=\"yes\">P. salmonis<\/jats:italic>\n                      genomes available shows that the\n                      <jats:italic toggle=\"yes\">comEC<\/jats:italic>\n                      gene\n                      <jats:italic toggle=\"yes\">,<\/jats:italic>\n                      encoding the DNA uptake channel required for natural transformation (NT) in competent bacteria, is interrupted by various transposable elements, whereas other NT determinants (\n                      <jats:italic toggle=\"yes\">comEA<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">comFB<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">comM<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">comL\/bamD<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">dprA<\/jats:italic>\n                      ) remain intact and highly conserved. Using\n                      <jats:italic toggle=\"yes\">P. salmonis<\/jats:italic>\n                      Psal-103 (EM-genogroup) and Psal-104b (LF-genogroup) as representatives of the two most prevalent genogroups in Chile, we combined comparative genomics, gene expression analysis, heterologous expression of\n                      <jats:italic toggle=\"yes\">comEC<\/jats:italic>\n                      using homologs from naturally competent bacteria, and CRISPRi-mediated knockdown of the remaining NT-related genes to examine their functionality. According to our results, expression of\n                      <jats:italic toggle=\"yes\">comEC<\/jats:italic>\n                      with homologs from\n                      <jats:italic toggle=\"yes\">Legionella pneumophila<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">Vibrio cholerae<\/jats:italic>\n                      impaired\n                      <jats:italic toggle=\"yes\">P. salmonis<\/jats:italic>\n                      viability in axenic culture, indicating a fitness burden. CRISPRi-mediated repression of\n                      <jats:italic toggle=\"yes\">comL\/bamD<\/jats:italic>\n                      revealed its essentiality in Psal-104b and a critical role during infection of the SHK-1 cell line in Psal-103. Repression of\n                      <jats:italic toggle=\"yes\">comM<\/jats:italic>\n                      tended to reduce cytopathogenicity in both strains, while repression of\n                      <jats:italic toggle=\"yes\">comFB<\/jats:italic>\n                      was associated with a modest delay in cytopathic damage in Psal-104b. Repression of\n                      <jats:italic toggle=\"yes\">recA<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">comEA<\/jats:italic>\n                      caused moderate reductions in cytopathic activity in Psal-103. By contrast,\n                      <jats:italic toggle=\"yes\">dprA<\/jats:italic>\n                      showed no detectable phenotypes. Together, our results indicate that despite the irreversible interruption of\n                      <jats:italic toggle=\"yes\">comEC<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">P. salmonis<\/jats:italic>\n                      has retained NT-related genes that contribute to fitness in a gene- and genogroup-specific manner, providing a framework to investigate how determinants of horizontal gene transfer function beyond DNA uptake.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Despite its major impact on salmon aquaculture,\n                        <jats:italic toggle=\"yes\">Piscirickettsia salmonis<\/jats:italic>\n                        remains poorly characterized at the functional level, largely due to long-standing limitations in genetic tractability. Here, we implement and combine multiple genetic approaches, including CRISPR interference, site-specific chromosomal integration, and heterologous gene expression, to functionally interrogate natural transformation (NT)-related genes in this pathogen. Using representative strains from the two most prevalent Chilean genogroups, we show that conserved NT-associated genes contribute to bacterial physiology and infection in a genogroup-dependent manner. Moreover, and beyond the specific biological findings, this work establishes a versatile genetic platform for functional studies in\n                        <jats:italic toggle=\"yes\">P. salmonis<\/jats:italic>\n                        , expanding the experimental toolbox available to study this pathogen and supporting future efforts aimed at understanding its biology and the development of novel biotechnological approaches.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.03173-25","type":"journal-article","created":{"date-parts":[[2026,2,4]],"date-time":"2026-02-04T14:01:02Z","timestamp":1770213662000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Genomic and functional dissection of natural transformation-related genes in\n                    <i>Piscirickettsia salmonis<\/i>"],"prefix":"10.1128","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-0181-2846","authenticated-orcid":true,"given":"Sebasti\u00e1n","family":"Higuera-Llant\u00e9n","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02cafbr77","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratorio de Gen\u00e9tica e Inmunolog\u00eda Molecular, Facultad de Ciencias, Instituto de Biolog\u00eda, Pontificia Universidad Cat\u00f3lica de Valpara\u00edso, Campus Curauma","place":["Valpara\u00edso, Chile"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8533-6942","authenticated-orcid":false,"given":"Nicolas","family":"Ojeda","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02cafbr77","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratorio de Gen\u00e9tica e Inmunolog\u00eda Molecular, Facultad de Ciencias, Instituto de Biolog\u00eda, Pontificia Universidad Cat\u00f3lica de Valpara\u00edso, Campus Curauma","place":["Valpara\u00edso, Chile"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Javiera","family":"Protz","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02cafbr77","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratorio de Gen\u00e9tica e Inmunolog\u00eda Molecular, Facultad de Ciencias, Instituto de Biolog\u00eda, Pontificia Universidad Cat\u00f3lica de Valpara\u00edso, Campus Curauma","place":["Valpara\u00edso, Chile"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0438-8252","authenticated-orcid":false,"given":"Sergio H.","family":"Marshall","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02cafbr77","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratorio de Gen\u00e9tica e Inmunolog\u00eda Molecular, Facultad de Ciencias, Instituto de Biolog\u00eda, Pontificia Universidad Cat\u00f3lica de Valpara\u00edso, Campus Curauma","place":["Valpara\u00edso, Chile"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.3147\/jsfp.25.107"},{"key":"e_1_3_3_3_2","first-page":"407","article-title":"British Columbia. 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