CBET-EPSRC: Developing standardized cell-free systems for prototyping complex synthetic biology circuits and pathways

这项研究是响应NSF工程- UKRI工程和物理科学研究理事会机会NSF 18-067的资助。存在设计和实现细胞内功能电路和反应途径的能力。该项目利用无细胞系统来克服与活细胞中这些活动相关的一些困难。测试细胞内性能的必要性将可能的途径限制在对宿主细胞无毒的那些途径。不幸的是，无细胞系统的性能已被证明是不稳定的，表现出较差的可靠性。该项目将建立一个可靠的，可重复的，无细胞的平台。如果能够在无细胞环境中完成，设计-构建-测试周期可以大大缩短。这将对科学研究、工程设备和STEM教育产生有益的影响。作为与伦敦帝国理工学院的合作项目，该项目还将促进研究交流，为美国学生提供国际研究经验。该项目的主要目标是标准化工程E.大肠杆菌和其他微生物。 这些系统将用于探索无细胞原型的边界，并使人们能够更详细地了解关键机制。这可以加速无细胞系统在工业和学术界的使用和更广泛的应用。 长期愿景是建立无细胞系统作为实施合成生物回路，途径和系统的平台，其中模块化和复杂的生物分子系统可以以系统的方式进行工程设计。 该项目旨在通过实验表征和计算建模的结合来克服无细胞系统的一些当前限制。 如果成功，该项目将把无细胞系统的实用性扩展到更广泛的电路和通路。 该项目的具体目标是：（1）开发易于理解的无细胞反应系统，适用于各种细胞的原型电路和途径;（2）使用无细胞提取物对复杂合成生物学组件和核心机制进行表征和建模;以及（3）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，被认为值得支持。

This research was funded in response to the NSF Engineering - UKRI Engineering and Physical Sciences Research Council Opportunity NSF 18-067. The capability exists to design and implement functioning circuits and reaction pathways inside cells. This project makes use of cell-free systems to overcome some of the difficulties associated with these activities in living cells. The necessity to test performance inside cells limits the possible pathways to those that are not toxic to the host cell. Unfortunately, the performance of cell-free systems has proven to be erratic, exhibiting poor reliability. This project will establish a reliable, reproducible, cell-free platform. The design-build-test cycle could be shortened dramatically if it could be accomplished in a cell-free environment. This would have a beneficial impact on scientific research, engineered devices, and STEM education. As a collaboration with Imperial College London, this project will also promote research exchanges, providing US students with international research experiences. The primary goal of this project is to standardize cell-free systems from engineered E. coli and other organisms. Such systems will be used to explore the boundaries of cell-free prototyping and enable more detailed understanding of key mechanisms. This could accelerate the usage and broader utility of cell-free systems in industry and academia. The long term vision is to establish cell-free systems as a platform for implementation of synthetic biological circuits, pathways, and systems, where modular and complex biomolecular systems can be engineered in a systematic fashion. This project seeks to overcome some of the current limitations of cell-free systems through a combination of experimental characterization and computational modeling. If successful, this project will expand the utility of cell-free systems to a broader class of circuits and pathways. The specific objectives of the project are (1) the development of well-understood cell-free reaction systems suitable for prototyping circuits and pathways for a variety of cells; (2) characterization and modeling of complex synthetic biology components and core mechanisms using cell-free extracts; and (3) development of new biochemical indicator modules for monitoring and characterizing circuit performance.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.