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Cell-free gene expression in synthetic vesicles

A bottom-up approach is used to build an artificial cell as a programmable phospholipid vesicle. A cell-free expression system is encapsulated in phospholipid vesicles. The translation machinery is used as the hardware and the DNA as the software. In vitro transcription and translation is maintained a few days in cell-sized vesicles with the internal expression of membrane channel alpha-hemolysin. This toxin allows exchange of nutrients and removal of byproducts with an external feeding solution. To transform the synthetic membrane into an active interface, amphipathic peptides, insertion mechanism of integral membrane proteins and lipid composition have been tested. Specific degradation mechanisms of the synthesized messengers and proteins are introduced to create a sink and get a homeostatic state. Perspectives and limitations of this approach will be discussed.