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

Author

Listed:
  • Alicia Broto

    (Imperial College London)

  • Erika Gaspari

    (Wageningen University & Research
    European & Developing Countries Clinical Trials Partnership (EDCTP))

  • Samuel Miravet-Verde

    (The Barcelona Institute of Science and Technology, Dr Aiguader 88)

  • Vitor A. P. Martins Santos

    (Wageningen University & Research
    LifeGlimmer GmbH)

  • Mark Isalan

    (Imperial College London)

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.

Suggested Citation

  • Alicia Broto & Erika Gaspari & Samuel Miravet-Verde & Vitor A. P. Martins Santos & Mark Isalan, 2022. "A genetic toolkit and gene switches to limit Mycoplasma growth for biosafety applications," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-29574-0
    DOI: 10.1038/s41467-022-29574-0
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    References listed on IDEAS

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    1. Elitza Deltcheva & Krzysztof Chylinski & Cynthia M. Sharma & Karine Gonzales & Yanjie Chao & Zaid A. Pirzada & Maria R. Eckert & Jörg Vogel & Emmanuelle Charpentier, 2011. "CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III," Nature, Nature, vol. 471(7340), pages 602-607, March.
    2. Eva Yus & Jae-Seong Yang & Adrià Sogues & Luis Serrano, 2017. "A reporter system coupled with high-throughput sequencing unveils key bacterial transcription and translation determinants," Nature Communications, Nature, vol. 8(1), pages 1-12, December.
    3. Marie Trussart & Eva Yus & Sira Martinez & Davide Baù & Yuhei O. Tahara & Thomas Pengo & Michael Widjaja & Simon Kretschmer & Jim Swoger & Steven Djordjevic & Lynne Turnbull & Cynthia Whitchurch & Mak, 2017. "Defined chromosome structure in the genome-reduced bacterium Mycoplasma pneumoniae," Nature Communications, Nature, vol. 8(1), pages 1-13, April.
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