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

FMRG：生物：DNA

• 批准号: 2134772
• 负责人: Elisa Franco
• 金额: $ 300万
• 依托单位: University of California-Los Angeles
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2134772&HistoricalAwards=false
• 关键词: FMRG; Bio; DNA; & RNA

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.

空间组织、分离和分类的能力是任何先进制造工艺的关键。虽然这些任务已经在自上而下的工业生化过程中掌握，但将这些操作嵌入微米和纳米尺度的生化反应器中仍然具有挑战性。微型反应器可用于在低体积、低成本反应中提取、分类、分离和组织组分，并且对于稀缺、有毒或高价值配体特别相关。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，灵感来自细胞凝聚现象，将小分子，核酸，蛋白质和整个反应途径分隔开来。 通过开发构建DNA和RNA浓缩物的技术蓝图，这项研究将促进在生物技术，制药和化学工程中采用这种强大的方法，并有助于产生新的劳动力。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，加州大学旧金山分校和加州理工学院进行测试，并将向公众提供。这种方法利用了核酸的热力学和动力学特性，使生物浓缩物的创新技术系统化。该项目旨在：（i）将DNA和RNA浓缩物发展成为未来的制造技术;（ii）吸引大学生参与，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;以及（iii）扩大国内制造高价值生物分子的能力。基于DNA和RNA纳米技术的进步，该项目将贡献DNA和RNA单体库，这些单体浓缩成液滴，这些液滴含有与分离和生产相关的特定分子和途径。实验将由可定制的主机冷凝物设计的预测模型指导，并通过设计-构建-测试管道将展示与传感，药物制造和医疗保健相关的可扩展分子，细胞和途径的液体分离。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。由于核酸天然存在于活的生物体中，因此可以将定制人工浓缩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分子和细胞生物科学司，工程理事会化学、生物医学、环境和运输系统司，教育和人力资源理事会本科教育司，以及数学和物理科学理事会的化学部。该奖项反映了NSF的法定使命，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，

项目成果

Chemical reaction motifs driving non-equilibrium behaviours in phase separating materials

驱动相分离材料非平衡行为的化学反应基序

• DOI: 10.1098/rsif.2023.0117
• 发表时间: 2023
• 期刊: Journal of The Royal Society Interface
• 影响因子: 3.9
• 作者: Osmanović, Dino;Franco, Elisa
• 通讯作者: Franco, Elisa

Light-controlled growth of DNA organelles in synthetic cells

• DOI: 10.1098/rsfs.2023.0017
• 发表时间: 2023-08-11
• 期刊: INTERFACE FOCUS
• 影响因子: 4.4
• 作者: Agarwal，Siddharth;Dizani，Mahdi;Franco，Elisa
• 通讯作者: Franco，Elisa

Robust Microphase Separation Through Chemical Reaction Networks

通过化学反应网络实现稳健的微相分离

• DOI: 10.1109/lcsys.2023.3283934
• 发表时间: 2023
• 期刊: IEEE Control Systems Letters
• 影响因子: 3
• 作者: Blanchini, Franco;Franco, Elisa;Giordano, Giulia;Osmanović, Dino
• 通讯作者: Osmanović, Dino