Design and construct modular transcriptional repressors to facilitate the development of living diagnostics

设计和构建模块化转录抑制子以促进活体诊断的发展

• 批准号: 9879608
• 负责人: Tsz Yan Clement Chan
• 金额: $ 39.17万
• 依托单位: UNIVERSITY OF NORTH TEXAS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9879608
• 关键词: Animals; Behavior; Bioinformatics; Biological; Biological Monitoring; Biomedical Engineering; Biosensor; Cell physiology; Cells; Chemicals; Clinical; Communities; Complex; Cues; DNA; DNA Binding; Data; Detection; Development; Devices; Diagnosis; Diagnostic; Engineering; Family; Gene Expression; Genetic; Goals; Health; Human; Hybrids; Ligands; Link; Medical; Mission; Monitor; Organism; Outcome; Output; Pathway interactions; Peptides; Performance; Physiological; Property; Protein Family; Proteins; Public Health; Regulation; Research; Scientist; Series; Signal Transduction; Signaling Molecule; Structure; System; Testing; Transcription Repressor; Transcriptional Regulation; Tweens; United States National Institutes of Health; Work; base; behavioral response; biological systems; cell type; cellular engineering; design; design and construction; flexibility; hybrid protein; in vivo; inducible gene expression; innovation; practical application; programs; promoter; response; signal processing; small molecule; synthetic biology; tool

PROJECT SUMMARY/ABSTRACT Developing multi-input, multi-output genetic circuits has to date been a significant challenge due to the fact that natural genetic systems are only capable to connect one single chemical input to one specific promoter to control gene expression, which poses a major barrier to 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 implement new cellular functions for practical applications. As important steps toward these goals, Aim #1 is to identify functional modules among TetR family homologs for small molecule sensing and DNA recognition. The central hypothesis is that TetR family repressors are composed of discrete and functional modules for detecting ligand molecules and for interacting with promoters to control gene expression, in which swapping these modules leads to hybrid repressors with new combinations of allosteric and DNA-binding properties. The strategy to achieve Aim #1 is to use bioinformatics approaches to analyze sequence and structure information of TetR homologs, predicting functional protein modules, and to experimentally validate the predictions by assessing the performance of these hybrid repressors during in vivo transcription regulation. For Aim #2, the research team proposes to create a genetic circuit platform that facilitates the use of various organisms for monitoring multiple physiological parameters. The working hypothesis is that modular repressors are able to serve as biosensors in many types of organisms to realize a genetic circuit design for simultaneous detection of multiple physiological changes. This aim is independent from Aim #1 because we can achieve the goal by using modular repressors developed previously from another protein family. The strategy to achieve Aim #2 is to engineer promoters in a range of organisms, in which the resulting promoters can be controlled by the desirable repressors. These engineered inducible expression systems can then be used to implement the circuit design for multiple signal detection. The contribution of this project is expected to be the establishment of a design principle for creating modular parts from TetR homologs and also, the creation of a genetic circuit platform for monitoring multiple physiological parameters by using various organisms. This contribution will be significant because it is expected to release many new possibilities in circuit topologies for engineering organisms for biomedical uses, including the biomonitoring platform developed in this project, which has a great potential to be used for diagnosis of biomedical conditions. Therefore, the proposed work is expected to move the field vertically at both the basic and applied levels.

项目摘要/摘要 迄今为止，由于事实 天然遗传系统仅能够将一个化学输入与一个特定启动子连接到 控制基因表达，这为创建具有复杂信号的工程生物带来了主要障碍 生物医学应用的响应行为。该研究团队的长期目标是建立 强大的策略来构建遗传回路的生物部分，并使用这些部分实施新的策略 用于实际应用的蜂窝功能。作为朝着这些目标的重要步骤，目标＃1是确定 小分子传感和DNA识别的TER家族同源物之间的功能模块。中央 假设是Tetr家族阻遏物由用于检测配体的离散和功能模块组成 分子并与启动子相互作用以控制基因表达，其中交换这些模块 导致具有变构和DNA结合特性的新组合的混合阻遏物。策略 实现目标＃1是使用生物信息学方法分析TERT的顺序和结构信息 同源物，预测功能蛋白模块，并通过评估实验验证预测 在体内转录调控过程中，这些杂交阻遏物的性能。对于AIM＃2，研究 团队建议创建一个遗传电路平台，以促进使用各种生物进行监测 多个生理参数。工作假设是模块化阻遏物能够充当 许多类型生物的生物传感器实现遗传回路设计，以同时检测多个 生理变化。这个目标独立于目标＃1，因为我们可以通过使用 模块化阻遏物以前是从另一个蛋白质家族开发的。实现目标2的策略是 一系列生物的工程师启动子可以由生物体控制。 理想的阻遏物。然后可以使用这些工程化的诱导表达系统来实施 多个信号检测的电路设计。预计该项目的贡献将是建立 设计原理是从Tetr同源物创建模块化部分的设计原理，还创建了遗传回路 通过使用各种生物来监测多个生理参数的平台。这个贡献将是 重要的是因为预计它将在工程上释放许多新的可能性 生物医学用途的生物，包括该项目中开发的生物监测平台 用于诊断生物医学条件的巨大潜力。因此，拟议的工作应 在基本水平和应用水平上垂直移动字段。