Synthetic P-bodies: Coupling gene expression and ribonucleoprotein granules in synthetic cell vesicles for sensing and response

合成生物学家正在努力构建具有特定功能和所需应用的合成细胞，例如根除引起感染的微生物的能力或分解环境中有毒化学物质的能力。 这些合成细胞需要的一个基本功能是主动感知和响应外部环境的能力。 该项目的目标是测试一种被称为可编程体或P体的合成区室化系统，该系统旨在为合成细胞提供感知和响应环境线索的能力。 除了合成P-体的发展，该项目将允许一个跨学科的团队来培训研究生和本科生，扩大知识和专业网络提供给参与者，并设计和提供一个学习模块“合成生命伦理”。该项目整合了自下而上合成生物学和内在无序蛋白质，核酸结合蛋白质和核酸的液-液相分离工程的进展。具体而言，目标是开发一种通用的合成细胞平台，其整合细胞内合成P体，其以类似于天然无膜细胞器的方式动态响应并调节基因表达。这些体包括相分离的核酸和固有无序蛋白质与核糖核蛋白的工程融合体。它们的动态和可逆的形成和溶解是由化学诱导的基因表达介导的，以实现化学传感。分子水平的响应同样能够通过合成P体的触发形成和溶解来螯合或释放分子货物。该项目将合成细胞整合到功能性杂化材料系统中，用于控制细菌生物膜形成的特定应用。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。

Synthetic biologists are embarking on efforts to build synthetic cells with specific functions and desired applications such as the ability to eradicate infection-causing microbes or the ability to breakdown toxic chemicals in the environment. One basic function these synthetic cells will need is the ability to actively sense and respond to the external environment. The goal of this project is to test a synthetic compartmentalization system known as programmable bodies or P-bodies, which is designed to provide synthetic cells with the ability to sense and respond to environmental cues. In addition to the development of synthetic P-bodies, this project will allow an interdisciplinary team to train graduate and undergraduate students, to expand the intellectual and professional networks available to participants, and to design and deliver a learning module on “synthetic life ethics.”This project integrates advances in bottom-up synthetic biology and engineering of liquid-liquid phase separation of intrinsically disordered proteins, nucleic-acid binding proteins, and nucleic acids. Specifically, the goal is to develop a versatile synthetic cell platform that integrates intracellular synthetic P-bodies which dynamically respond to, and regulate, gene expression in manners analogous to natural membraneless organelles. These bodies comprise phase-separated nucleic acids and engineered fusions of intrinsically disordered proteins and ribonucleoproteins. Their dynamic and reversible formation and dissolution is mediated by chemically induced gene expression to enable chemical sensing. Molecular-level response is likewise enabled by the triggered formation and dissolution of the synthetic P-bodies to sequester or release molecular cargo. The project integrates synthetic cells into functional hybrid materials systems for the specific application of controlling formation of bacterial biofilms.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.