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

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Manufacturers culture cells to produce vaccines and protein-based drugs. These products are complex and difficult to synthesize because they are composed of proteins. Growing cells is not very efficient for production because a cell uses a lot of energy and raw material to grow and maintain itself in addition to making a particular product. This limitation spurs an interest in creating synthetic cells to mimic the cellular processes that support product formation rather than cell growth. Some cellular processes occur in separate compartments to avoid interference with each other. This project will attempt to create synthetic compartments where biological reactions can take place. Success would make creating complex molecules and proteins easier. It would also increase our understanding of how cells perform and coordinate critical functions. The project results will be shared with the general public through videos. Lab tours will be provided to girls through the Career Day for Girls program at Northwestern. Underrepresented minority students from Chicago Public Schools will also be mentored. These efforts should encourage more students to pursue careers in STEM-related fields.Controlling when and where biomolecular reactions take place remains a significant challenge in the design of cell-free systems. In this project, the role of compartmentalization on a model biosynthetic reaction will be explored systematically. Compartments will be examined to uncover how spatial segregation and the nature of the segregating compartment can enhance cell-free synthesis. The approach will couple compartmentalization with techniques in cell-free metabolism and regulation. Computational modeling will be employed to modularly implement production of a model biomolecule. The contribution of the project results will be to markedly improve biological synthesis in cell-free environments by integrating compartmentalization, controlled transport, and cell-free expression systems to achieve greater temporal control and expand the environments in which such systems can be employed.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.