INSPIRE Track 1: Protocells as a Platform for Bottom-up Synthetic Biology

1344298Wang该INSPIRE奖的部分资金来自工程理事会CBET部门的生物技术、生化和生物质工程项目，生物科学理事会MCB部门的合成和系统生物学项目，数学和物理科学理事会DMR部门的生物材料项目，以及国际和综合活动办公室。在这个项目中，人造细胞将被设计成具有定义的构建块，这些构建块可能小到单个基因、蛋白质和RNA。实现这种方法的平台是原始细胞，这是一种通过定向自组装构建的纳米工程结构，提供了细胞内部和表面膜的独立定义。在这种自下而上的方法中，可以确切地知道单元格中有什么，因此，不受控制的变量被最小化。最初，将开发一个概念验证，其中包括膜和细胞内成分的信号通路，随着更多成分的加入，导致更大的复杂性。最终，一个具有基本功能的最小活细胞将被设计和研究。因此，本课题的三个主要目标是：(1)创建一个通用的合成生物学平台，研究信号转导和参数可控的蛋白质合成；(2)建立调控合成细胞内信号转导和蛋白质合成的机制模型；(3)弥合体外实验与细胞实验之间的差距。这些目标将通过设计和制造具有越来越复杂水平的连续版本的原始细胞来实现。通过在系统生物学和合成生物学的背景下整合纳米科学、分子生物传感器、成像和生物化学，将建立一个高度跨学科的平台来重建和模拟细胞的复杂性，为理解细胞环境中分子网络的设计原则提供一个通用和系统的方法。这项工作将对理解生物分子网络中的信息流和纳米尺度上的定向自组装具有重要意义。因此，该项目的成功将对生物纳米技术和仿生设备产生变革性影响。此外，这项研究将与教育工作相结合，以确保本科生和高中生，特别是代表性不足的群体有机会和鼓励从事尖端的跨学科研究。

1344298Wang This INSPIRE award is partially funded by the Biotechnology,Biochemical and Biomass Engineering Program in the CBET Division in the Directorate for Engineering, the Synthetic and Systems Biology Program in the Division of MCB in the Directorate for Biological Sciences, the Biomaterials Program in the Division of DMR in the Directorate for Mathematics and Physical Sciences, and by the Office of International and Integrative Activities.In this project artificial cells will be engineered with defined building blocks that may be as small as individual genes, proteins, and RNA. The platform that enables this approach is the protocell, a nanoengineered construct built by directed self-assembly that provides for independent definition of the cell interior and the surface membrane. In this bottom-up approach it is exactly known what is in the cell, and, therefore, uncontrolled variables are minimized. Initially a proof-of-concept will be developed in which a signaling pathway including both membrane and intracellular components is assembled, leading to greater complexity as more components are added. Ultimately a minimal living cell with basic functions will be engineered and studied. Accordingly, three major goals of the proposal are: (1) to create a general synthetic biology platform to study signal transduction and protein synthesis with controlled parameters; (2) to build mechanistic models governing the signal transduction and protein synthesis inside the synthesized cell; (3) to bridge the gap between in vitro and in cell experiments. These goals will be achieved by employing successive versions of protocells designed and fabricated with increasing levels of complexity.By integrating nanoscience, molecular biosensors, imaging, and biochemistry in the context of systems biology and synthetic biology, a highly interdisciplinary platform to reconstitute and model cellular complexity will be established providing a general and systematic approach to understand the design principles of molecular networks in a cellular environment. This work will have fundamental implications for understanding information flow in biomolecular networks and directed self-assembly at the nanoscale. As such, the success of this project will have transformative impact on bionanotechnology and biomimetic devices. In addition, the research will be integrated with educational efforts to ensure that undergraduate and high school students and particularly underrepresented groups have the opportunity and encouragement to work in cutting-edge interdisciplinary research.