Xu Xiaopei, Du Zhaoxia, Liu Renmei, Li Ting, Zhao Yuzheng, Chen Xianjun, Yang Yi
CAS Center for Excellence in Brain Science, Shanghai Institutes for Biological Sciences , Chinese Academy of Sciences , Shanghai 200031 , China.
ACS Synth Biol. 2018 Sep 21;7(9):2045-2053. doi: 10.1021/acssynbio.8b00180. Epub 2018 Sep 4.
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 Sic1 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.
光作为一种极具吸引力的促动器，能够对多种细胞活动进行时空控制。在本研究中，我们为酵母细胞开发了一种单组分光控基因表达系统，称为yLightOn系统。yLightOn系统不依赖外源辅助因子，具有超过500倍的开/关比、极低的渗漏率、快速的表达动力学以及高空间分辨率。我们通过分别严格控制His3和Sic1基因的表达，证明了yLightOn系统在调节细胞生长和细胞周期方面的实用性。此外，我们构建了一个双向表达模块，可通过光同时控制两个基因的表达。以ClpX和ClpP作为报告基因，观察到了带有ssrA标签的蛋白质的快速、定量且空间特异性降解。我们认为，这种单组分光遗传学系统将对理解细胞基因调控网络以及设计用于合成生物学的稳健遗传电路有极大帮助。
