Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago 8331150, Chile.
Millennium Institute for Integrative Biology (iBio), Santiago 8331150, Chile.
ACS Synth Biol. 2023 Jan 20;12(1):71-82. doi: 10.1021/acssynbio.2c00338. Epub 2022 Dec 19.
Cell communication is a widespread mechanism in biology, allowing the transmission of information about environmental conditions. In order to understand how cell communication modulates relevant biological processes such as survival, division, differentiation, and apoptosis, different synthetic systems based on chemical induction have been successfully developed. In this work, we coupled cell communication and optogenetics in the budding yeast . Our approach is based on two strains connected by the light-dependent production of α-factor pheromone in one cell type, which induces gene expression in the other type. After the individual characterization of the different variants of both strains, the optogenetic intercellular system was evaluated by combining the cells under contrasting illumination conditions. Using luciferase as a reporter gene, specific co-cultures at a 1:1 ratio displayed activation of the response upon constant blue light, which was not observed for the same cell mixtures grown in darkness. Then, the system was assessed at several dark/blue-light transitions, where the response level varies depending on the moment in which illumination was delivered. Furthermore, we observed that the amplitude of response can be tuned by modifying the initial ratio between both strains. Finally, the two-population system showed higher fold inductions in comparison with autonomous strains. Altogether, these results demonstrated that external light information is propagated through a diffusible signaling molecule to modulate gene expression in a synthetic system involving microbial cells, which will pave the road for studies allowing optogenetic control of population-level dynamics.
细胞通讯是生物学中一种广泛存在的机制，允许传递有关环境条件的信息。为了了解细胞通讯如何调节相关的生物学过程，如生存、分裂、分化和凋亡，已经成功开发了基于化学诱导的不同合成系统。在这项工作中，我们在 budding yeast 中结合了细胞通讯和光遗传学。我们的方法基于两种菌株的连接，通过在一种细胞类型中光依赖性产生α-因子信息素来诱导另一种类型的基因表达。在对两种菌株的不同变体进行单独表征后，通过在对比光照条件下组合细胞来评估光遗传细胞间系统。使用荧光素酶作为报告基因，在特定的共培养物中以 1:1 的比例显示出在持续蓝光下的响应激活，而在黑暗中生长的相同细胞混合物中则没有观察到这种情况。然后，在几个暗/蓝光转换中评估了该系统，其中响应水平取决于光照提供的时刻。此外，我们观察到通过修改两种菌株之间的初始比例可以调整响应幅度。最后，与自主菌株相比，双种群系统显示出更高的倍增值。总之，这些结果表明，外部光信息通过可扩散的信号分子传播，以调节涉及微生物细胞的合成系统中的基因表达，这将为允许光遗传控制群体水平动力学的研究铺平道路。
