Single-Cell Chemical Genetics Platform

单细胞化学遗传学平台

• 批准号: 7140628
• 负责人: CARL L HANSEN
• 金额: $ 13.18万
• 依托单位: UNIVERSITY OF BRITISH COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7140628
• 关键词: biological signal transduction; biomedical automation; cell cell interaction; cell sorting; charge coupled device camera; chemical genetics; microfluidics; microscopy; mitogen activated protein kinase; perfusion; photonics; single cell analysis; small molecule; technology /technique development; yeasts

DESCRIPTION (provided by applicant): Deciphering complex protein interactions and regulatory networks is of central interest in understanding cellular function and development, and in elucidating the mechanism of disease. To this end, chemical biology has emerged as a powerful technique for dissecting these networks through the controlled perturbation of protein function using biologically active small molecule compounds. However, conventional chemical biology studies provide only population averaged measurements of large number of cells. This ensemble averaging conceals the ever-present heterogeneity in cellular response, and obscures the underlying biology. Furthermore, current technologies provide only crude means of modulating the chemical environment over time, making them poorly suited to the study of kinetics and of response to sequential stimuli. Deciphering the complex underlying networks and cell-cell variations manifest in cellular response requires new technologies for the quantitative and time-resolved analysis of single cells under well-defined and time-varying chemical environments. We will combine state-of-the-art techniques in microfluidics and microscopy to build a single-cell chemical biology platform for conducting highly parallel studies of the response of single cells to time-varying sequences of chemical conditions and bioactive small molecules. This fully automated platform will advance the state-of-the-art in single cell analysis and should find broad applicability to the study of many cell types. This new technology will be validated in the quantitative analysis of cell-cell heterogeneity, kinetics, and robustness arising in the yeast pheromone/filamentous growth pathway which serves as an archetypical model for studying the highly conserved MARK signal transduction conduits. Relevance to public health: A new instrument will be built to enable the analysis of single cells exposed to precisely defined and time- varying chemicals conditions and drugs. By allowing for experimentation at the single cell level, this instrument will improve both our understanding of, and our ability to treat, disease. Furthermore, the ability to precisely manipulate and interrogate single cells will enable new lines of inquiry in relevant health fields including cancer research, stem cell research, and developmental biology.

描述（由申请人提供）：解密复杂的蛋白质相互作用和调节网络是理解细胞功能和发育以及阐明疾病机制的核心兴趣。为此，化学生物学已成为一种强大的技术，用于通过使用生物活性的小分子化合物对蛋白质功能的受控扰动来解剖这些网络。但是，传统的化学生物学研究仅提供人群平均测量大量细胞。这种合奏平均掩盖了细胞反应中永远存在的异质性，并掩盖了潜在的生物学。此外，当前的技术仅提供了随着时间的推移调节化学环境的粗略手段，使其适合研究动力学和对顺序刺激的反应。在细胞响应中，解释复杂的基础网络和细胞细胞变化需要新的技术，以对单个细胞进行定量和时间分解分析，并在定义明确的和时变的化学环境下进行定量和时间分解。我们将结合微流体和显微镜中的最新技术，以建立一个单细胞化学生物学平台，以对单个细胞对化学条件和生物活性小分子的时变序列的反应进行高度平行的研究。这个完全自动化的平台将在单细胞分析中推进最先进的平台，并应对许多细胞类型的研究发现广泛的适用性。这项新技术将在酵母网络/丝素/丝状生长途径中的细胞细胞异质性，动力学和鲁棒性的定量分析中得到验证，该途径是研究高度保守的标记信号传递导管的原型模型。 与公共卫生的相关性：将建立一种新工具，以便对暴露于精确定义和时间变化的化学物质条件和药物的单个单元进行分析。通过允许在单细胞水平进行实验，该仪器将提高我们对疾病的理解和治疗能力。此外，精确操纵和询问单细胞的能力将在包括癌症研究，干细胞研究和发育生物学在内的相关健康领域的新询问线。