Expanding the synthetic cell toolkit: programmable fusion for complex genetic circuits and a synthetic cell cycle

Abstract

Synthetic biology as a scientific discipline is relatively new; however, major strides have been taken to develop current in vitro technologies into platforms that can imitate life. Currently, synthetic cell technologies often address only one feature of life (cell division, protein expression, DNA replication, etc.) due to the technological limitations of the current systems. In this dissertation, major improvements in the synthetic cell field are achieved by coupling well-established liposomal technologies with other novel technologies. The first of these is the development of a synthetic cell system that uses programmable liposomal fusion events to control and regulate complex genetic circuits originating from within synthetic cells. The second combines several synthetic cell technologies to realize a model cell cycle within these life-like liposomal bioreactors. Although the product of this dissertation is quite obviously not fully “alive,” major improvements in current methodologies were achieved, allowing for these artificially produced synthetic cells to blur the line a even more between in vitro systems and biological life.