Development of Genetic Sensors and Circuits for Creating Novel Cellular Behaviors

项目摘要/摘要 即使合成生物学取得了最近的进展，但在开发遗传电路方面仍然是一个重大挑战 这涉及到多个输入和输出。这是因为自然遗传系统只能 将一个单一的化学输入连接到一个特定的启动子，以控制基因的表达。这构成了一个 为生物医学创造具有复杂信号响应行为的工程化生物的重大障碍 申请。这个研究团队的长期目标是建立健全的战略来构建 基因电路的生物部分，并利用这些部分来扩大研究人员设计新的 生物医学应用中的细胞功能。在他们最近的进展中，该团队建立了一个模块交换 Laci和TetR家族中的调节器构建遗传传感器的策略，他们利用了这些 设计传感器以开发几种新的遗传电路。这项研究的两个方向 代表着朝着团队未来五年的长期目标迈出的重要一步。第一个方向是 提高转录调节器作为模块化生物传感器的工程能力。具体地说，团队 计划1)建立修改调节器的设计原则，以增强其作为生物传感器的性能 以及2)将模块交换应用于广泛的调节器系列。中心假设是，每一个 一个家族中的调节子包含一个配体结合模块(LBM)和一个DNA结合模块(DBM)，用于 分别用于检测输入信号和用于与启动子交互的目的；如果关键模块-模块 保持交互，来自不同监管机构的LBM和DBM可以混合和匹配以创建 混合调节器，具有输入传感和DNA识别特性的新组合。为了他们的第二次 在这个方向上，该团队建议利用混合调节器来探索各种生物体中的新电路设计， 旨在满足生物医学领域的新需求。这一努力包括开发蜂窝设备以 持续和同时监测一系列有毒污染物，这提供了一种手段来评估 摄入在受污染的食物和水中常见的有毒物质。作为一名早期调查员， PI和他的团队已经在两个拟议的方向上取得了重大进展，表明 他们有很高的资格从事拟议的项目。预计该项目的贡献将是 建立用于从许多家族的调节器创建模块化零件的设计原则 遗传电路设计和实现方面的进展。这一贡献将是重大的，因为它是 预计将在生物医学用途的电路拓扑中发布许多新的可能性，包括监控 将在此程序中创建的设备。总体方法是创新的，因为它代表了一种新的 使用蛋白质工程和细胞工程方法来提高公众健康和安全的方法。 因此，拟议的工作预计将在科学和社会层面产生积极影响。

PROJECT SUMMARY/ABSTRACT Even with recent advances in synthetic biology, it remains a major challenge in developing genetic circuits that involve multiple inputs and outputs. This is because natural genetic systems are only capable of connecting one single chemical input to one specific promoter to control gene expression. This poses a significant barrier in creating engineered organisms with complex signal response behavior for biomedical applications. The long-term goals of this research team are to establish robust strategies for constructing biological parts of genetic circuits, and to use these parts to expand researchers’ ability in engineering new cellular functions for biomedical applications. In their recent progress, the team established a module swapping strategy for building genetic sensors from regulators in the LacI and TetR families and they harnessed these engineered sensors to develop several novel genetic circuits. The two directions in this proposed research represent important steps toward the team’s long-term goals in the next five years. The first direction is to advance the capabilities in engineering transcriptional regulators as modular biosensors. Specifically, the team plans to 1) establish design principles of modifying regulators for enhancing their performance as biosensors and 2) apply module swapping to a wide range of regulator families. The central hypothesis is that each regulator within a family contains a ligand-binding module (LBM) and a DNA-binding module (DBM) for the purpose of detecting an input signal and for interacting with a promoter, respectively; if key module-module interactions are maintained, LBMs and DBMs from different regulators can be mixed and matched to create hybrid regulators with new combinations of input sensing and DNA recognition properties. For their second direction, the team proposes to harness hybrid regulators to explore novel circuit designs in various organisms, aiming to meet emerging needs in biomedical fields. This effort includes developing cellular devices to continuously and simultaneously monitor a range of toxic pollutants, which provides a means to assess the intake of toxicants that are commonly found in contaminated food and water. As an Early Stage Investigator, the PI and his team have already generated significant progress on both proposed directions, showing that they are highly qualified to pursue the proposed projects. The contribution of this project is expected to be the establishment of design principles for creating modular parts from regulators in many families and the advancement in genetic circuit design and implementation. This contribution will be significant because it is expected to release many new possibilities in circuit topologies for biomedical uses, including monitoring devices that will be created in this program. The overall approach is innovative because it represents a new way of using protein engineering and cellular engineering approaches to enhance public health and safety. Therefore, the proposed work is expected to generate positive impacts at both scientific and societal levels.