Assembly and disassembly of artificial cells

The work in this thesis was part of Artificial Cells with Distributed Cores, ACDC, a European funded project. The aim of the project is to emulate the structure and dynamics of living cells and cellular tissues by incorporating biochemical and biophysical systems capable of sensing, reconfiguration, synthesis and biochemical energy production - all essential processes of living cells - built from the bottom-up approach. As the first steps towards that future vision, the aim is to produce an artificial technological construct and process that recapitulates some aspects of living systems on the microscale, and therefore this project will produce exemplars of secondary living technologies. Here, I focused on studying the dynamic interactions between artificial cells through the assembly and disassembly of cores taking advantage of a technology based on DNA as a linker molecule between cores. A novel system was designed to be controlled and responsive to external environmental inputs as a basis for a future technology that will harness it for several applications, including sensing and actuation in the environments for bioremediation and detection and rebalancing of any imbalances at the blood level. In this thesis I was able to create a system where artificial cells emulsion droplets (EDs) and giant unilamellar vesicles (GUVs) were produced and can assemble when different populations are decorated with a specific DNA sequence. Disassembly of artificial cells was investigated and a system prototype was produced. GUVs were generated capable of in situ transcription, so that the RNA produced inside can diffuse out upon addition of a pore forming protein on the surface of vesicles membranes. This RNA can act as a reactive molecule able to disturb DNA assembly of vesicles and produce a disassembled state. The system described here is the foundation for a future portable system that forms the basis for a portable technology that can respond to external stimuli by providing clear, highly visible outputs and whose applications will be many.