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

1344298Wang这个INSPIRE奖的部分资金来自工程局CBET部门的生物技术、生化和生物量工程计划，生物科学局MCB部门的合成和系统生物学计划，数学和物理科学委员会DMR部门的生物材料计划，以及国际和综合活动办公室。在这个项目中，人工细胞将用定义的构建块进行工程，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，它提供了对细胞内部和表面膜的独立定义。在这种自下而上的方法中，单元格中的内容是确切知道的，因此，不受控制的变量被最小化。最初，将开发一种概念验证，其中包括膜和细胞内组件的信号通路被组装，随着添加更多组件，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，该提案的三个主要目标是：(1)建立一个通用的合成生物学平台，研究信号转导和蛋白质合成，并控制参数；(2)建立控制合成细胞内信号转导和蛋白质合成的机制模型；(3)弥合体外和细胞实验之间的差距。通过在系统生物学和合成生物学的背景下集成纳米科学、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重建和模拟细胞复杂性的平台，为理解细胞环境中分子网络的设计原理提供一种通用和系统的方法。这项工作将对理解生物分子网络中的信息流和纳米尺度的定向自组装具有基本意义。因此，该项目的成功将对生物纳米技术和仿生设备产生革命性的影响。此外，研究将与教育努力相结合，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前沿跨学科研究。

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.