Department of Life Sciences, Imperial College London, London, SW7 2AZ, UK.
Laboratory of Systems and Synthetic Biology, Wageningen University & Research, Wageningen, the Netherlands.
Nat Commun. 2022 Apr 7;13(1):1910. doi: 10.1038/s41467-022-29574-0.
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.
支原体具有极其精简的基因组，并能很好地适应其众多宿主，从宿主那里获取必要的营养。由于其相对简单的基因组，支原体已被用于开发生物技术应用的底盘。然而，缺乏用于基因组操作和严格调控的强大而精确的工具包，阻碍了任何实质性的进展。在此，我们描述了一种用于肺炎支原体的强大遗传工具包的构建，并成功地将其用于设计合成基因开关，以控制和限制支原体生长，用于生物安全控制应用。我们发现，这些合成基因回路在最小细胞环境中具有长期的稳定性和鲁棒性。通过这项工作，我们为开发可行且强大的生物安全系统奠定了基础，以利用合成支原体底盘作为治疗应用的活减毒载体。
