Light is a highly attractive actuator that allows spatiotemporal control of diverse cellular activities. In this study, we developed a single-component light-switchable gene expression system for yeast cells, termed yLightOn system. The yLightOn system is independent of exogenous cofactors, and exhibits more than a 500-fold ON/OFF ratio, extremely low leakage, fast expression kinetics, and high spatial resolution. We demonstrated the usefulness of the yLightOn system in regulating cell growth and cell cycle by stringently controlling the expression of His3 and ^ΔNSic1 genes, respectively. Furthermore, we engineered a bidirectional expression module that allows the simultaneous control of the expression of two genes by light. With ClpX and ClpP as the reporters, the fast, quantitative, and spatially specific degradation of ssrA-tagged protein was observed. We suggest that this single-component optogenetic system will be immensely helpful in understanding cellular gene regulatory networks and in the design of robust genetic circuits for synthetic biology.

中文翻译：

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单组分光遗传学系统允许严格转换酵母细胞中的基因表达。

光是一种极具吸引力的执行器，它可以对各种细胞活动进行时空控制。在这项研究中，我们开发了一种用于酵母细胞的单组分光开关基因表达系统，称为yLightOn系统。yLightOn系统独立于外源辅因子，并具有超过500倍的开/关比，极低的泄漏，快速的表达动力学和高的空间分辨率。我们证实在通过严格控制的表达调节细胞生长和细胞周期的yLightOn系统的有用性HIS3和^ΔN SIC1基因分别。此外，我们设计了一种双向表达模块，该模块允许通过光同时控制两个基因的表达。以ClpX和ClpP为报告基因，观察到了ssrA标签蛋白的快速，定量和空间特异性降解。我们建议，这种单组分光遗传学系统将对理解细胞基因调控网络和设计用于合成生物学的稳健遗传电路有极大的帮助。