Key Laboratory of Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
ACS Synth Biol. 2020 Aug 21;9(8):2144-2153. doi: 10.1021/acssynbio.0c00211. Epub 2020 Jul 15.
Cell-free systems, as part of the synthetic biology field, have become a critical platform in biological studies. However, there is a lack of research into developing a switch for a dynamical control of the transcriptional and translational process. The optogenetic tool has been widely proven as an ideal control switch for protein synthesis due to its nontoxicity and excellent time-space conversion. Hence, in this study, a blue light-regulated two-component system named YF1/FixJ was incorporated into an based cell-free system to control protein synthesis. The corresponding cell-free system successfully achieved a 5-fold dynamic protein expression by blue light repression and 3-fold dynamic expression by blue light activation. With the aim of expanding the applications of cell-free synthetic biology, the cell-free blue light-sensing system was used to perform imaging, light-controlled antibody synthesis, and light-triggered artificial cell assembly. This study can provide a guide for further research into the field of cell-free optical sensing. Moreover, it will also promote the development of cell-free synthetic biology and optogenetics through applying the cell-free optical sensing system to synthetic biology education, biopharmaceutical research, and artificial cell construction.
无细胞系统作为合成生物学领域的一部分，已成为生物学研究的关键平台。然而，在开发用于转录和翻译过程动态控制的开关方面，相关研究还比较缺乏。由于其非毒性和优异的时空转换特性，光遗传学工具已被广泛证明是蛋白质合成的理想控制开关。因此，在本研究中，我们将一种名为 YF1/FixJ 的蓝光调控双组分系统整合到基于无细胞的系统中，以控制蛋白质合成。相应的无细胞系统成功地实现了蓝光抑制时 5 倍的动态蛋白表达和蓝光激活时 3 倍的动态表达。为了拓展无细胞合成生物学的应用，我们使用无细胞蓝光感应系统进行了成像、光控抗体合成以及光触发人工细胞组装。本研究可为无细胞光学感应领域的进一步研究提供指导。此外，通过将无细胞光学感应系统应用于合成生物学教育、生物制药研究和人工细胞构建，也将促进无细胞合成生物学和光遗传学的发展。
