Optogenetic Tools for Control of Public Goods in Saccharomyces cerevisiae.
Optogenetic Tools for Control of Public Goods in Saccharomyces cerevisiae.
复制标题
DOI：
10.1128/msphere.00581-21
复制
发表时间：
2021-08-25
期刊：
影响因子：
4.8
通讯作者：
McClean MN
中科院分区：
文献类型：
--
作者：
Moreno Morales N;Patel MT;Stewart CJ;Sweeney K;McClean MN
微生物生活在密集且多样化的群落中，细胞之间的相互作用引导群落的发展和表型。在空间和时间上干扰特定细胞间相互作用的能力为确定微生物群落的关键相互作用和设计规则提供了强大的途径。使用光遗传学工具来调节这些相互作用的方法提供了希望，因为光可以在空间和时间上得到精确控制。我们报告了新的质粒，用于将光遗传学系统快速整合到酿酒酵母中，以设计感兴趣基因表达的光控制。在原理验证研究中，我们证明了控制模型合作相互作用的能力，即转化酶（SUC2）的表达，它允许酿酒酵母水解蔗糖并将其用作碳源。我们证明，可以通过调制光强度和照明的空间控制来在空间和时间上调整这种协作相互作用的强度。光的空间控制允许合作者和作弊者在空间上隔离，我们表明空间中合作和抑制相互作用之间的相互作用可以导致模式形成。我们的策略可用于实现对酿酒酵母中感兴趣的基因表达的时空控制，以扰乱细胞间和种间相互作用。重要性微生物生态学的最新进展强调了细胞间相互作用在控制微生物群落的发育、组成和恢复力方面的重要性。为了更好地理解这些相互作用在管理社区发展中的作用，能够以受控的方式改变它们至关重要。光遗传学控制的相互作用比静态扰动或化学控制的相互作用具有优势，因为可以在空间和时间上操纵光，并且不需要添加营养物或抗生素。在这里，我们报告了一种快速实现对重要模式生物酿酒酵母中感兴趣基因的光控制的系统，并证明通过控制转化酶的表达，我们可以控制协同相互作用。该方法将有助于理解含有酿酒酵母的天然和合成微生物群落中的细胞间和种间相互作用，并可作为在其他群落中实施该方法的原理证明。
Microorganisms live in dense and diverse communities, with interactions between cells guiding community development and phenotype. The ability to perturb specific intercellular interactions in space and time provides a powerful route to determining the critical interactions and design rules for microbial communities. Approaches using optogenetic tools to modulate these interactions offer promise, as light can be exquisitely controlled in space and time. We report new plasmids for rapid integration of an optogenetic system into Saccharomyces cerevisiae to engineer light control of expression of a gene of interest. In a proof-of-principle study, we demonstrate the ability to control a model cooperative interaction, namely, the expression of the enzyme invertase (SUC2) which allows S. cerevisiae to hydrolyze sucrose and utilize it as a carbon source. We demonstrate that the strength of this cooperative interaction can be tuned in space and time by modulating light intensity and through spatial control of illumination. Spatial control of light allows cooperators and cheaters to be spatially segregated, and we show that the interplay between cooperative and inhibitory interactions in space can lead to pattern formation. Our strategy can be applied to achieve spatiotemporal control of expression of a gene of interest in S. cerevisiae to perturb both intercellular and interspecies interactions. IMPORTANCE Recent advances in microbial ecology have highlighted the importance of intercellular interactions in controlling the development, composition, and resilience of microbial communities. In order to better understand the role of these interactions in governing community development, it is critical to be able to alter them in a controlled manner. Optogenetically controlled interactions offer advantages over static perturbations or chemically controlled interactions, as light can be manipulated in space and time and does not require the addition of nutrients or antibiotics. Here, we report a system for rapidly achieving light control of a gene of interest in the important model organism Saccharomyces cerevisiae and demonstrate that by controlling expression of the enzyme invertase, we can control cooperative interactions. This approach will be useful for understanding intercellular and interspecies interactions in natural and synthetic microbial consortia containing S. cerevisiae and serves as a proof of principle for implementing this approach in other consortia.