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

空间组织、分离和分类的能力是任何先进制造过程的关键。虽然这些任务已经在自上而下的工业生化过程中掌握，但将这些操作嵌入到微纳米尺度的生化反应器中仍然具有挑战性。微观反应器用于在低体积、低成本的反应中提取、分类、分离和组织组分，尤其适用于稀有、有毒或高价值的配体。该项目将建立设计工具和组件，以构建由DNA和RNA制成的微观液体反应器，从分隔小分子、核酸、蛋白质和整个反应途径的细胞凝聚现象中获得灵感。通过开发构建DNA和RNA凝聚体的技术蓝图，本研究将促进这种强大方法在生物技术、制药和化学工程领域的应用，并为新一代劳动力做出贡献。为了实现这一目标，本科生将参与研究，研究将被整合到教育模块中，这些模块将在帕萨迪纳城市学院、加州大学洛杉矶分校、加州大学圣迭戈分校和加州理工学院进行测试，并将向公众开放。这种方法充分利用了核酸的热力学和动力学特性，使生物凝析油的创新技术系统化。该项目旨在：(i)将DNA和RNA凝聚物发展成为未来的制造技术；（ii）吸引本科生，开发和传播教育工具，为积极参与这一新兴技术的劳动力做好准备；（三）扩大国内高价值生物分子的制造能力。基于DNA和RNA纳米技术的进步，该项目将提供DNA和RNA单体文库，这些文库可以凝聚成液滴，其中包含与分离和生产相关的特定分子和途径。实验将以设计可定制宿主凝析物的预测模型为指导，并通过设计-构建-测试管道展示可扩展的分子、细胞和与传感、药物制造和医疗保健相关的途径的液体分离。pi团队包括DNA自组装系统（Rothemund），工程DNA和RNA反应网络（Franco），核酸系统生物物理学（Fygenson）和生物化学教育（Blatti）领域的领导者。由于核酸自然存在于生物体中，因此可以将定制人工冷凝物的操作与细胞和组织的操作无缝集成。该项目由生物科学理事会的分子和细胞生物科学司、工程理事会的化学、生物医学、环境和运输系统司、教育和人力资源理事会的本科教育司以及数学和物理科学理事会的化学司共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.