Synthetic biology-based detection of micronutrients with minimal equipment

使用最少的设备进行基于合成生物学的微量营养素检测

• 批准号: 9383810
• 负责人: Mark Philip-Walter Styczynski
• 金额: $ 35.51万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-21 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9383810
• 关键词: Address; Affect; Area; Bacteria; Behavior; Binding; Binding Proteins; Biological Assay; Biological Models; Biological Preservation; Biosensor; Blood; Blood Tests; Blood specimen; Cell Culture Techniques; Cell Survival; Cell physiology; Cells; Centrifugation; Cessation of life; Chelating Agents; Child; Child Mortality; Color; Custom; Data; Detection; Development; Diagnosis; Diagnostic; Dietary Intervention; Electricity; Engineering; Environment; Epidemiologist; Equipment; Escherichia coli; Freeze Drying; Future; Genetic; Goals; Growth; Health; Hour; Human; Hybrids; Immunologic Factors; Impairment; Incidence; Intervention; Laboratories; Location; Logistics; Malaria; Malnutrition; Measurement; Measures; Methods; Micronutrients; Minerals; Nucleic Acids; Nutrient; Nutritional; Nutritional status; Oranges; Output; Pathway interactions; Patients; Performance; Pigments; Pneumonia; Policy Making; Population; Populations at Risk; Production; Research; Resources; Sampling; Series; Serum; Site; Source; Techniques; Technology; Testing; Time; Transportation; Vitamins; Work; Zinc; Zinc deficiency; base; burden of illness; cost; epidemiology study; extracellular; improved; metabolic engineering; mortality; novel; novel strategies; nutrition; personalized medicine; point of care; portability; promoter; response; sensor; success; synthetic biology; transcription factor

Project Summary The goal of the proposed work is to create bacteria that detect the level of a micronutrient (vitamin or mineral) in a sample, with ultimate application to human blood tests. The micronutrient to be detected is zinc. These cells will enable the later development of minimal-equipment testing of blood zinc levels in remote, resource-poor locations, a significant improvement in both the time and cost necessary using current state-of-the-art methods. The cells use a genetic circuit controlled by zinc-sensitive transcription factors to produce different colored pigment based on the zinc concentration in the sample, indicating whether zinc levels are acceptable or low. Thus, the cells function as a sort of easy-to-read “bacterial litmus test”. Strong preliminary work supports the likelihood of our success. The first aim involves creating the bacterial strain capable of producing three different pigments in response to different levels of extracellular zinc. The second aim involves making the existing circuit repress pigment production until the assay is performed, allowing for the pre-culture of colorless cells that will enable fast coloration and overcome obstacles to sensor cell survival in human blood samples.The third aim consists of tuning the cells' performance when grown in actual blood samples, since work to date has used laboratory growth medium which may yield different results than growth in the long-term target samples. This project will yield the underlying technology that can be used for the first-ever bacterial blood test for low-resource settings to provide population-level assessment of micronutrient status. By being low-cost and essentially point-of-care, such a long-term result would enable nutritional epidemiologists and policymakers to make more informed decisions about nutritional interventions, as well as to assess the success of interventions after the fact, potentially improving the health of millions of undernourished people. Moreover, the pigment-producing genetic circuit establishes a framework that can be applied for the development of other micronutrient sensors using different nutrient-binding proteins, potentially allowing a whole panel of inexpensive tests.

项目摘要 这项工作的目标是创造出一种能够检测微量营养素（维生素）水平的细菌 或矿物质），最终应用于人类血液测试。微量营养素 检测到锌。这些单元将使以后开发的最低限度的设备测试， 血液锌水平在偏远，资源贫乏的地区，一个显着的改善，在这两个时间 和使用现有技术方法所需的成本。这些细胞利用基因回路 由锌敏感转录因子控制，产生不同颜色的色素， 样品中的锌浓度，表明锌水平是否可接受或低。因此，在本发明中， 这些细胞起到了一种易于阅读的“细菌石蕊测试”的作用。强大的前期工作支持 我们成功的可能性。第一个目标是创造一种能够 产生三种不同的色素来响应不同水平的细胞外锌。第二 Aim涉及使现有的电路抑制颜料产生直到进行化验， 允许预培养无色细胞，这将能够快速着色并克服 第三个目标包括调整人类血液样本中的传感器细胞存活的障碍。 细胞在实际血液样本中生长时的性能，因为迄今为止的工作一直使用实验室 生长培养基，其可能产生与长期目标样品中的生长不同的结果。 该项目将产生可用于有史以来第一次细菌血液的基础技术 在低资源环境下进行测试，以提供人口一级的微量营养素状况评估。通过 由于成本低，基本上是即时护理，这样的长期结果将使营养 流行病学家和政策制定者做出更明智的决定， 干预措施，以及评估干预措施的成功后，事实上，可能改善 数百万营养不良者的健康。此外，产生色素的基因电路 建立了一个框架，可用于开发其他微量营养素传感器 使用不同的营养素结合蛋白，可能允许一整套廉价的测试。