Broto, AliciaGaspari, ErikaMiravet Verde, Samuel, 1992-Martins dos Santos, VitorIsalan, Mark2022-05-312022-05-312022Broto A, Gaspari E, Miravet-Verde S, Martins Dos Santos VAP, Isalan M. A genetic toolkit and gene switches to limit Mycoplasma growth for biosafety applications. Nat Commun. 2022 Apr 7;13(1):1910. DOI:10.1038/s41467-022-29574-02041-1723http://hdl.handle.net/10230/53326Mycoplasmas have exceptionally streamlined genomes and are strongly adapted to their many hosts, which provide them with essential nutrients. Owing to their relative genomic simplicity, Mycoplasmas have been used to develop chassis for biotechnological applications. However, the dearth of robust and precise toolkits for genomic manipulation and tight regulation has hindered any substantial advance. Herein we describe the construction of a robust genetic toolkit for M. pneumoniae, and its successful deployment to engineer synthetic gene switches that control and limit Mycoplasma growth, for biosafety containment applications. We found these synthetic gene circuits to be stable and robust in the long-term, in the context of a minimal cell. With this work, we lay a foundation to develop viable and robust biosafety systems to exploit a synthetic Mycoplasma chassis for live attenuated vectors for therapeutic applications.application/pdfeng© Alicia Broto et al. 2022. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were madeGenèticaGenòmicaMicoplasmosisA genetic toolkit and gene switches to limit Mycoplasma growth for biosafety applicationsinfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1038/s41467-022-29574-0info:eu-repo/semantics/openAccess