{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T01:18:44Z","timestamp":1790212724655,"version":"4.1.0"},"reference-count":29,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2024,9,18]],"date-time":"2024-09-18T00:00:00Z","timestamp":1726617600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/doi.org\/10.1128\/ASMCopyrightv2"},{"start":{"date-parts":[[2024,9,18]],"date-time":"2024-09-18T00:00:00Z","timestamp":1726617600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"crossref","award":["ANR-19-CE14-0045-002"],"award-info":[{"award-number":["ANR-19-CE14-0045-002"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2024,9,18]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              The efficient natural transformation of\n              <jats:italic>Neisseria meningitidis<\/jats:italic>\n              allows the rapid construction of bacterial mutants in which the genes of interest are interrupted or replaced by antibiotic-resistance cassettes. However, this proved to be a double-edged sword, i.e., although facilitating the genetic characterization of this important human pathogen, it has limited the development of strategies for constructing markerless mutants without antibiotic-resistance markers. In addition, efficient tools for complementation or labeling are also lacking in\n              <jats:italic>N. meningitidis<\/jats:italic>\n              . In this study, we significantly expand the meningococcal genetic toolbox by developing new and efficient tools for the construction of markerless mutants (using a dual counterselection strategy), genetic complementation (using integrative vectors), and cell labeling (using a self-labeling protein tag). This expanded toolbox paves the way for more in-depth genetic characterization of\n              <jats:italic>N. meningitidis<\/jats:italic>\n              and might also be useful in other\n              <jats:italic>Neisseria<\/jats:italic>\n              species.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                <jats:italic>Neisseria meningitidis<\/jats:italic>\n                and\n                <jats:italic>Neisseria gonorrhoeae<\/jats:italic>\n                are two important human pathogens. Research focusing on these bacteria requires genetic engineering, which is facilitated by their natural ability to undergo transformation. However, the ease of mutant engineering has led the\n                <jats:italic>Neisseria<\/jats:italic>\n                community to neglect the development of more sophisticated tools for gene editing, particularly for\n                <jats:italic>N. meningitidis<\/jats:italic>\n                . In this study, we have significantly expanded the meningococcal genetic toolbox by developing novel and efficient tools for markerless mutant construction, genetic complementation, and cell tagging. This expanded toolbox paves the way for more in-depth genetic characterization of\n                <jats:italic>N. meningitidis<\/jats:italic>\n                and might also be useful in other\n                <jats:italic>Neisseria<\/jats:italic>\n                species.\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/aem.00880-24","type":"journal-article","created":{"date-parts":[[2024,8,14]],"date-time":"2024-08-14T13:01:35Z","timestamp":1723640495000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":5,"title":["Expanding the genetic toolbox for\n            <i>Neisseria meningitidis<\/i>\n            with efficient tools for unmarked gene editing, complementation, and labeling"],"prefix":"10.1128","volume":"90","author":[{"given":"Morgane","family":"Wuckelt","sequence":"first","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, UFR de M\u00e9decine, Paris, France"},{"name":"Inserm U1151, Institut Necker-Enfants Malades, CNRS UMR 8253, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Audrey","family":"Laurent","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, UFR de M\u00e9decine, Paris, France"},{"name":"Inserm U1151, Institut Necker-Enfants Malades, CNRS UMR 8253, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cl\u00e9mence","family":"Mouville","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, UFR de M\u00e9decine, Paris, France"},{"name":"Inserm U1151, Institut Necker-Enfants Malades, CNRS UMR 8253, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie","family":"Meyer","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, UFR de M\u00e9decine, Paris, France"},{"name":"Inserm U1151, Institut Necker-Enfants Malades, CNRS UMR 8253, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne","family":"Jamet","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, UFR de M\u00e9decine, Paris, France"},{"name":"Inserm U1151, Institut Necker-Enfants Malades, CNRS UMR 8253, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Herv\u00e9","family":"Lecuyer","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, UFR de M\u00e9decine, Paris, France"},{"name":"Inserm U1151, Institut Necker-Enfants Malades, CNRS UMR 8253, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xavier","family":"Nassif","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Paris Cit\u00e9, UFR de M\u00e9decine, Paris, France"},{"name":"Inserm U1151, Institut Necker-Enfants Malades, CNRS UMR 8253, Paris, 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