RoL: RAISE: DESYN-C3: A platform for Modular Pseudo-Organelles for Compartmentalization and Control of Pseudo-Cell Processes

Spatial organization is critical to life, yet it remains conspicuously absent from most engineered systems, particularly synthetic cells or pseudo-cells. This challenge arises from the difficulty of maintaining spatial segregationwhile permitting controlled molecular exchange between reaction pathways.Thechallenge of combining structural organization with chemical communication must be solved in order to generate pseudo-cell platforms that can support complex cellular functions required for technological applications and fundamental studies. Our long-term goal is to develop a model pseudo-cell chassis capable of supporting the modular implementation of biosensing, biocomputing, and biomanufacturing processes in the form of compartmentalized enzymatic reactions and biochemical circuits. Towards this goal, our objective for this RAISE is to develop a platform in which we can precisely control when and where distinct biochemical reactions take place, and to use modeling and experimental techniques to analyze and optimize intended reactions. Using techniques in cell-free metabolism and regulation, compartmentalization strategies, and computational modeling, we will design and analyze nanoscale compartments we term pseudo-organelles in order to modularly implement a biochemical reaction. As a result, we will uncover rules to combine structural organization with chemical communication in order to generate pseudo-cell platforms that can support complex cellular functions required for technological applications and fundamental studies.