Development of Genetic Sensors and Circuits for Creating Novel Cellular Behaviors

项目总结/摘要 即使合成生物学取得了最新的进展，它仍然是开发遗传电路的一个主要挑战 涉及多个输入和输出。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表达。这会带来一定的 在创造具有复杂信号响应行为的生物医学工程生物体方面的重大障碍 应用.该研究小组的长期目标是建立强有力的战略， 基因电路的生物部分，并利用这些部分来扩大研究人员的能力，在工程新的 生物医学应用的细胞功能。在他们最近的进展中，该团队建立了一个模块交换 从LacI和TetR家族的调节因子中构建遗传传感器的策略， 设计传感器来开发几种新的基因电路。本研究的两个方向 代表了团队在未来五年内实现长期目标的重要步骤。第一个方向是 推进作为模块化生物传感器的工程化转录调节器的能力。具体来说，团队 计划1）建立修改调节器的设计原则，以提高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。核心假设是， 一个家族内的调节子包含用于调节的配体结合模块（LBM）和DNA结合模块（DBM）。 分别用于检测输入信号和用于与启动子相互作用的目的; 保持相互作用，来自不同监管机构的LBM和DBM可以混合和匹配， 具有输入传感和DNA识别特性的新组合的混合调节器。为他们的第二 方向，该团队提出利用混合调节器来探索各种生物体中的新颖电路设计， 旨在满足生物医学领域的新兴需求。这项工作包括开发蜂窝设备， 连续和同时监测一系列有毒污染物，这提供了一种评估 摄入了在受污染的食物和水中常见的有毒物质。作为一个早期的研究者， PI和他的团队已经在这两个建议的方向上取得了重大进展，表明 他们完全有资格推行建议的项目。预计该项目的贡献将是 建立设计原则，用于从许多系列的调节器中创建模块化部件， 基因电路设计和实现的进展。这一贡献将是重大的，因为它是 预计将释放许多新的可能性，在电路拓扑结构的生物医学用途，包括监测 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 利用蛋白质工程和细胞工程方法来提高公共健康和安全。 因此，预计拟议的工作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.