Design Considerations for Synthetic Cells

Efforts to understand life as we know it and life as it can be have culminated in the field of synthetic cell research, which aims to build life from the bottom up using individual biological components. Recent progress in the field has enabled the reconstitution of many functions of living cells in synthetic cells, from cell-cell communication to membrane protein expression and function. However, future progress in the field is limited by many challenges, including irreproducibility, lack of predictability, difficulties in integrating existing synthetic cell modules (or subsystems), and the need for autonomous functionalities. In this work, I describe my efforts towards addressing these challenges. In Chapter 2, I describe sources of variability in transcription-translation (TX-TL) systems, the biological machinery used to implement biomolecular programs in synthetic cells. In Chapter 3, I describe a novel methodology for readily building more predictive models of TX-TL performance. In Chapter 4, I present a design for a proof-of-concept for integrating an energy regeneration subsystem and a motility subsystem to achieve autonomous programmable motility and highlight some early successes towards achieving that goal. Throughout this work, I highlight many design principles for building synthetic cells reproducibly, more predictably, and with novel functionalities.