(174bs) Bringing Physical Signals into Cell-Free Expression

The cell-free protein synthesis (CFPS) system is a typical protein production platform in the field of synthetic biology. However, there are limitations in controlling protein synthesis in the CFPS system. Compared with the traditional method of chemical stimulus, physical signal is an ideal switch to achieve precise spatiotemporal control with few side effects. For cell-free system, physical signal is a kind of ideal spatiotemporal control approach to replace chemical substances, realizing high accuracy with little side effect. Here we designed the methods of using physical signals including light and temperature to control gene expression in cell-free systems. A blue light-regulated two-component system named YF1/FixJ and a temperature-controlled cell-free protein synthesis (tcCFPS) system containing cI repressor based on E. coli was carried out. Moreover, the artificial cell controlled by blue light and temperature was constructed to expand the applications of physical signal controlled cell-free system. This study provides a new effective method for active protein synthesis in a cell-free system and a potential means of drug synthesis and delivery.