(176c) Driving Encapsulated Biological Reactions with DNA-Functionalized Vesicles

Controlling biological reactions spatially and temporally remains a significant challenge in the design of cell-mimetic systems. Cellular systems utilize extracellular vesicles, like exosomes and ectosomes, to transport biological molecules to target cells and initiate biochemical reactions. Recapitulating this behavior in synthetic vesicles is a promising approach to selectively control interactions between vesicle compartments and would enable the initiation of biological reactions in complex aqueous environments. DNA strands tethered to phospholipid vesicles, which act as SNARE protein mimetics, drive membrane fusion. Inspired by viral membrane fusion, we explore how membrane phase-segregation alters DNA-mediated fusion dynamics. We show that membrane phase-segregation provides an energetic driver for membrane fusion that increases the efficiency of DNA-mediated fusion events. Using this system, we show that orthogonality provided by DNA tethers allows us to direct fusion and delivery of DNA cargo to specific vesicle populations. We then demonstrate that vesicle fusion between DNA-tethered vesicles can be used to deliver plasmids into vesicles containing in vitro protein expression machinery and leads to the synthesis of soluble and membrane proteins. We expect that this method to engineer orthogonal fusion between DNA-tethered vesicles will provide a new strategy to control the spatio-temporal dynamics of cell-free reactions, and create opportunities to engineer complex artificial cellular systems.