FMRG: Bio: DNA & RNA Condensate Droplets for Programmable Separation and Manufacture of Biomolecules

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The ability to spatially organize, separate, and sort is key to any advanced manufacturing process. While these tasks have been mastered in top-down industrial biochemical processes, it remains challenging to embed these operations in biochemical reactors at the micro- and nano-scale. Microscopic reactors are useful to extract, sort, separate, and organize components in low-volume, low-cost reactions, and are particularly relevant for scarce, toxic, or high-value ligands. This project will establish design tools and components to build microscopic liquid reactors made with DNA and RNA, taking inspiration from cellular condensation phenomena that compartmentalize small molecules, nucleic acids, proteins, and entire reaction pathways. By developing a technological blueprint to build DNA and RNA condensates, this research will promote the adoption of this powerful approach in biotechnology, pharmaceutics, and chemical engineering, and contribute to the generation of a new workforce. Toward this goal, undergraduates will be involved in the research and the research will be integrated in educational modules that will be tested at Pasadena City College, UCLA, UCSB, and Caltech, and will be made available to the general public. This approach takes advantage of the well-understood thermodynamic and kinetic properties of nucleic acids to systematize the innovative technology of biological condensates. The project aims to: (i) develop DNA and RNA condensates into a future manufacturing technology; (ii) engage undergraduates and develop and disseminate educational tools for preparing a workforce to actively participate in this emerging technology; and (iii) expand the capabilities for domestic manufacturing of high-value biomolecules. Building on advances in DNA and RNA nanotechnology, the project will contribute libraries of DNA and RNA monomers that condense into liquid droplets that host specific molecules and pathways that are relevant for separation and production. Experiments will be guided by predictive models for design of customizable host condensates, and through a design-build-test pipeline will demonstrate liquid separation of an expandable set of molecules, cells, and pathways that are relevant for sensing, drug manufacturing, and healthcare. The team of PIs includes leaders in the field of DNA self-assembling systems (Rothemund), engineered DNA and RNA reaction networks (Franco), biophysics of nucleic acid systems (Fygenson), and biochemistry education (Blatti). Because nucleic acids are naturally present in living organisms, it will be possible to seamlessly integrate the operation of the custom artificial condensates with that of cells and tissues. This project is jointly funded by the Division of Molecular and Cellular Biosciences in the Biological Sciences Directorate, the Division of Chemical, Biomedical, Environmental and Transport Systems in the Engineering Directorate, the Division of Undergraduate Education in the Education and Human Resources Directorate, and the Division of Chemistry in the Mathematical and Physical Sciences Directorate.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.