Design of a genetically programmable artificial cell system for biocatalysis

用于生物催化的基因可编程人工细胞系统的设计

• 批准号: 1916030
• 负责人: Claudia Schmidt-Dannert
• 金额: $ 62.46万
• 依托单位: University of Minnesota-Twin Cities
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1916030&HistoricalAwards=false
• 关键词: Design; genetically; programmable; artificial; cell

The machinery that drives protein synthesis can be purified intact from cells. Using this machinery, protein can be produced at the direction of synthetic DNA outside of a cell. This opens up a wide range of opportunities. Cell activities can be isolated and studied. Artificial systems can be designed to perform a limited number of cell functions. A cell-like system could be fabricated that produces a single chemical or pharmaceutical. The goal of this project is to encase cell-sized compartments inside polymer membranes. The compartments will contain protein synthesis machinery and DNA. The enzymes that are created will produce valuable chemicals from simple starting materials. DNA for different cell systems will be mixed and matched to allow rapid prototyping of artificial cell configurations. The project will provide training and mentoring opportunities for high-school, undergraduate and graduate students. STEM outreach activities aimed at PreK-12 students will be pursued using resources available locally.Advances in cell-free systems are enabling the bottom-up design of synthetic cells. These cells could be comprised of a genetically encoded multi-enzyme cascade. Spatial organization could be designed to mimic cellular environments. Rapid prototyping of biocatalytic cascades is a key capability of these systems. Cell-like systems would be specifically designed to meet biocatalytic process requirements. These would include robustness, operational stability and catalytic efficiency of enzymes, and membrane porosity for co-factor, substrate and product transport. The design of such, genetically programmable artificial cell-like systems for in vitro cascade biocatalysis has not yet been accomplished and poses challenges that this project will address. A framework will be developed for the design, fabrication and operation of a robust artificial cell system for multi-enzyme cascade catalysis. An industrially relevant redox cascade will constitute the model system. A robust chassis will be built with a composite membrane, enforced by a genetically programmed protein-based cell-wall that contain pores/channels that facilitate membrane transport. Biocatalysts will be expressed inside the compartments along with protein based macromolecular structures for spatial organization of enzymes and control microenvironments. Knowledge gained in this project will serve as a framework for the bottom-up design of a synthetic cell chassis with genetically programmable subcellular features and membrane properties. The artificial cells developed in this project will allow experimental testing and further development of mathematical models proposed for compartmentalized systems. Results from these studies will advance our understanding of biological systems and provide guidance for the rational design of synthetic cells for biomanufacturing and other applications.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.

驱动蛋白质合成的机器可以从细胞中完整地纯化出来。使用这种机制，蛋白质可以在细胞外合成DNA的方向上产生。这带来了广泛的机会。可以分离和研究细胞活动。人工系统可以被设计成执行有限数量的细胞功能。可以制造一种细胞样系统，产生单一的化学品或药物。该项目的目标是将细胞大小的隔室包裹在聚合物膜内。隔室将包含蛋白质合成机器和DNA。所产生的酶将从简单的起始材料中产生有价值的化学物质。不同细胞系统的DNA将被混合和匹配，以允许人工细胞配置的快速原型。该项目将为高中生、本科生和研究生提供培训和指导机会。将利用当地现有资源开展针对PreK-12学生的STEM外联活动。无细胞系统的进步使自下而上设计合成细胞成为可能。这些细胞可以由遗传编码的多酶级联组成。 空间组织可以被设计成模仿细胞环境。生物催化级联的快速原型是这些系统的关键能力。细胞样系统将被专门设计以满足生物催化过程的要求。这些将包括酶的稳健性、操作稳定性和催化效率，以及用于辅因子、底物和产物运输的膜孔隙率。这种用于体外级联生物催化的遗传可编程人工细胞样系统的设计尚未完成，并提出了本项目将解决的挑战。将开发一个框架的设计，制造和操作的一个强大的人工细胞系统的多酶级联催化。 一个工业相关的氧化还原级联将构成模型系统。一个强大的底盘将建立一个复合膜，加强了基因编程的蛋白质为基础的细胞壁，含有孔/通道，促进膜运输。生物催化剂将在隔室内沿着与基于蛋白质的大分子结构一起表达，用于酶的空间组织和控制微环境。在这个项目中获得的知识将作为一个框架，自下而上的设计与遗传可编程的亚细胞功能和膜特性的合成细胞底盘。在这个项目中开发的人工细胞将允许实验测试和进一步发展的数学模型提出的分隔系统。这些研究的结果将促进我们对生物系统的理解，并为生物制造和其他应用的合成细胞的合理设计提供指导。该奖项反映了NSF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。