{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T23:29:01Z","timestamp":1790724541641,"version":"4.1.0"},"reference-count":43,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2024,7,24]],"date-time":"2024-07-24T00:00:00Z","timestamp":1721779200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/doi.org\/10.1128\/ASMCopyrightv2"},{"start":{"date-parts":[[2024,7,24]],"date-time":"2024-07-24T00:00:00Z","timestamp":1721779200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"Canadian Government | Canadian Institutes of Health Research"},{"DOI":"10.13039\/100010318","name":"University of Manitoba","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100010318","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100010318","name":"University of Manitoba","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100010318","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100008794","name":"Research Manitoba","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100008794","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100010318","name":"University of Manitoba","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100010318","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100010318","name":"University of Manitoba","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100010318","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000082","name":"Cystic Fibrosis Canada","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100000082","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Canadian Government | Canadian Institutes of Health Research"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2024,7,24]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              Genome editing in non-model bacteria is important to understand gene-to-function links that may differ from those of model microorganisms. Although species of the\n              <jats:italic>Burkholderia cepacia<\/jats:italic>\n              complex (Bcc) have great biotechnological capacities, the limited genetic tools available to understand and mitigate their pathogenic potential hamper their utilization in industrial applications. To broaden the genetic tools available for Bcc species, we developed RhaCAST, a targeted DNA insertion platform based on a CRISPR-associated transposase driven by a rhamnose-inducible promoter. We demonstrated the utility of the system for targeted insertional mutagenesis in the Bcc strains\n              <jats:italic>B. cenocepacia<\/jats:italic>\n              K56-2 and\n              <jats:italic>Burkholderia multivorans<\/jats:italic>\n              ATCC17616. We showed that the RhaCAST system can be used for loss- and gain-of-function applications. Importantly, the selection marker could be excised and reused to allow iterative genetic manipulation. The RhaCAST system is faster, easier, and more adaptable than previous insertional mutagenesis tools available for Bcc species and may be used to disrupt pathogenicity elements and insert relevant genetic modules, enabling Bcc biotechnological applications.\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>\n              Species of the\n              <jats:italic>Burkholderia cepacia<\/jats:italic>\n              complex (Bcc) have great biotechnological potential but are also opportunistic pathogens. Genetic manipulation of Bcc species is necessary to understand gene-to-function links. However, limited genetic tools are available to manipulate Bcc, hindering our understanding of their pathogenic traits and their potential in biotechnological applications. We developed a genetic tool based on CRISPR-associated transposase to increase the genetic tools available for Bcc species. The genetic tool we developed in this study can be used for loss and gain of function in Bcc species. The significance of our work is in expanding currently available tools to manipulate Bcc.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/aem.00699-24","type":"journal-article","created":{"date-parts":[[2024,6,13]],"date-time":"2024-06-13T13:01:39Z","timestamp":1718283699000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":14,"title":["A CRISPR-Cas-associated transposon system for genome editing in\n            <i>Burkholderia cepacia<\/i>\n            complex species"],"prefix":"10.1128","volume":"90","author":[{"given":"Zhong Ling","family":"Yap","sequence":"first","affiliation":[{"name":"Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A. S. M. 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