The development of environmentally responsive synthetic biology systems

Over recent years, there has been a growing interest in the field of artificial cell development, as they offer tremendous potentials across many fields of science and technology. A significant amount of progress has been made, particularly in the field of bottom-up artificial cells, due to the advancement in synthetic biology which has allowed us to assemble artificial cells from biological components systematically. The ability to sense and respond to their environment, in particular, has been the centre of much attention as it plays a key role in many important biological processes such as controlled release, signalling and communication between cells. However, most of the developments to date are largely limited to simple structures with a single function due to technological limitations. Areas such as artificial tissues which required high structural complexity and collective functionality have been largely unexplored. This thesis details the development of environment responsive artificial cells as well as tissue mimics in the form of water in oil droplets and aims to address these lack of platform technologies. We developed an optofluidic platform technology DROPLAY, capable of generating functional droplet networks with user-defined 2D structures. Instead of assembling the droplet network mechanically, a stem-cell-like approach was used. By incorporating a light-responsive mechanism into droplets, we have induced droplet differentiation to generate functional droplet networks using a laser-based setup. The results from this technology provide the foundation for future development of artificial tissues or other complex membrane structures. In addition, numerous light-responsive mechanisms, suitable for the construction of artificial cells, both chemical and biological have been investigated. In particular, we have showed and enhanced Bacteriorhodopsin (BR) activity in the acidification of lipid vesicles. Furthermore, a light-induced calcium controlled release system has been established and shown great potential. These light-responsive mechanisms can be incorporated in DROPLAY technology in the future or be introduced to other membrane structures for constructing more complex environment sensing artificial cells.Open Acces