Development of Genetic Sensors and Circuits for Creating Novel Cellular Behaviors

开发用于创造新细胞行为的遗传传感器和电路

• 批准号: 10705649
• 负责人: Tsz Yan Clement Chan
• 金额: $ 36.01万
• 依托单位: UNIVERSITY OF NORTH TEXAS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705649
• 关键词: Address; Biological; Biosensor; Cell physiology; Chemicals; Complex; DNA; DNA Binding; Development; Devices; Engineering; Family; Food Contamination; Gene Expression; Genetic; Genetic Transcription; Goals; Health; Hybrids; Intake; Ligand Binding; Medical; Mission; Monitor; Organism; Outcome; Output; Performance; Property; Protein Engineering; Protein Family; Public Health; Qualifying; Research; Research Personnel; Safety; Signal Transduction; System; United States National Institutes of Health; Water; Work; behavioral response; cell behavior; cellular engineering; design; innovation; interdisciplinary approach; monitoring device; novel; pollutant; programs; promoter; sensor; synthetic biology; tool; toxicant

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.

项目摘要/摘要 即使在合成生物学方面取得了最新进展，这仍然是发展遗传回路的主要挑战 这涉及多个输入和输出。这是因为自然遗传系统只能 将一个化学输入与一个特定启动子连接到控制基因表达。这定位a 生物医学具有复杂信号响应行为的工程生物的重大障碍 申请。该研究团队的长期目标是建立构建强大的策略 遗传回路的生物部分，并使用这些零件来扩大研究人员在工程中的新能力 生物医学应用的细胞功能。在他们最近的进度中，团队建立了一个模块交换 从LACI和Tetr家族中的监管机构构建遗传传感器的策略，并利用这些 设计的传感器以开发几个新型的遗传回路。这项拟议研究的两个方向 代表着未来五年实现团队长期目标的重要步骤。第一个方向是 提高工程转录调节器作为模块化生物传感器的功能。具体来说，团队 计划1）建立修改监管机构以增强其作为生物传感器的性能的设计原则 2）将模块交换应用于广泛的监管家族。中心假设是每个 家庭中的调节器包含一个配体结合模块（LBM）和一个DNA结合模块（DBM） 分别检测输入信号并分别与启动子相互作用的目的；如果密钥模块模块 保持交互作用，可以将来自不同调节器的LBM和DBM混合并匹配以创建 具有输入灵敏度和DNA识别特性的新组合的混合调节剂。他们的第二个 方向，团队提议利用混合监管机构探索各种生物体的新型电路设计， 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，简单地监测一系列有毒污染物，这提供了一种评估的手段 在受污染的食物和水中常见的毒物摄入量。作为早期的调查员 PI和他的团队已经在两个建议的指示上都取得了重大进展，表明 他们有资格从事拟议的项目。预计该项目的贡献将是 建立设计原则，以创建许多家庭监管机构的模块化零件以及 遗传电路设计和实施的进步。这项贡献将是重要的，因为它是 预计将在生物医学用途的电路拓扑中释放许多新的可能性，包括监视 将在此程序中创建的设备。总体方法是创新的，因为它代表了一个新的 使用蛋白质工程和蜂窝工程方法来增强公共健康和安全的方法。 因此，预计拟议的工作将在科学和社会水平上产生积极的影响。