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得到掌握，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。微型反应器可用于提取、分类、分离和组织小体积、低成本反应中的组分，尤其适用于稀缺、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 制成的微型液体反应器，其灵感来自分隔小分子、核酸、蛋白质和整个反应途径的细胞凝聚现象。 通过开发构建 DNA 和 RNA 凝聚物的技术蓝图，这项研究将促进这种强大方法在生物技术、制药和化学工程中的采用，并为新一代劳动力的产生做出贡献。为了实现这一目标，本科生将参与研究，研究结果将整合到教育模块中，这些模块将在帕萨迪纳城市学院、加州大学洛杉矶分校、加州大学圣巴巴拉分校和加州理工学院进行测试，并向公众开放。这种方法利用了人们熟知的核酸热力学和动力学特性，将生物浓缩物的创新技术系统化。该项目旨在：(i) 将 DNA 和 RNA 凝聚物开发为未来的制造技术； (ii) 让本科生参与并开发和传播教育工具，以培养劳动力积极参与这一新兴技术； (iii) 扩大国内高价值生物分子的制造能力。该项目以 DNA 和 RNA 纳米技术的进步为基础，将提供 DNA 和 RNA 单体库，这些单体库可凝结成液滴，其中包含与分离和生产相关的特定分子和途径。实验将以预测模型为指导，用于设计可定制的主体冷凝物，并通过设计构建测试管道来演示与传感、药物制造和医疗保健相关的一组可扩展分子、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。该项目由生物科学局分子和细胞生物科学处、工程局化学、生物医学、环境和运输系统处、教育和人力资源局本科教育处和数学和物理科学理事会化学部。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

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