A genetic toolkit and gene switches to limit Mycoplasma growth for biosafety applications

Alicia Broto, Erika Gaspari, Samuel Miravet-Verde, Vitor A.P. Martins dos Santos, Mark Isalan*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Mycoplasmas 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.

Original languageEnglish
Article number1910
JournalNature Communications
Volume13
Issue number1
DOIs
Publication statusPublished - 7 Apr 2022

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