Nanotechnology for Systems Biology of Neural Stem Cells

神经干细胞系统生物学纳米技术

• 批准号: 7156954
• 负责人: LIAOHAI CHEN
• 金额: $ 51.29万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-20 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7156954
• 关键词: Arts; Atomic Force Microscopy; Bacteriophages; Binding; Biological Assay; Cell Proliferation; Cell Separation; Cells; Complementary DNA; Detection; Development; Disadvantaged; Ensure; Enzyme-Linked Immunosorbent Assay; Fluorescence; Fluorescence Spectroscopy; Generations; Goals; Immobilization; Individual; Libraries; Life; Measures; Membrane Proteins; Methods; Microscopy; Molecular; Nanotechnology; Needles; Noise; Pan Genus; Phage Display; Plasma; Play; Process; Protein-Protein Interaction Map; Proteins; Research; Role; Role playing therapy; Screening procedure; Solutions; Specificity; Spectrum Analysis; Surface; Systems Biology; Technology; United States National Institutes of Health; Ursidae Family; base; cDNA Library; concept; nano; nanofabrication; nerve stem cell; novel; particle; programs; protein structure function; single molecule; tool

DESCRIPTION (provided by applicant): The proposed research harnesses the opportunities provided by single molecule spectroscopy and micro/nano fabrication technologies to create an enabling tool for the systems biology research of neural stem cell. Its goal is to quickly map protein-protein interactions of the proteins, which may play a role in controlling the proliferation and differentiation for the neural stem cells. Its objective is to generate/identify all possible interacting partners for surface proteins of a neural stem cell and to confirm their functionalities in living cells. Its approach is to utilize single-phage-display to screen a cDNA library of neural stem cell for the isolation of the corresponding interaction partners. Single-phage-display (SPD) engages a unique combination of micro-fabrication, phage display and single molecule detection and characterization technology such as fluorescence correlation spectroscopy (FCS). The concept is based on 1) the compartmentalization of a library solution (by either micro-channels or micro-droplets) to a level that each compartment only hosts a single phage particle statistically; 2) the interrogation of binding status of the phage particle in each individual compartment by FCS and 3) the identification of the phage particle that binds to the target by phage display. To ensure the capability of identifying interaction partners "on demand", single phage display without FCS was also proposed as a backup approach.

描述（由申请人提供）：拟议的研究利用单分子光谱学和微/纳米制造技术提供的机会，为神经干细胞的系统生物学研究创建一个有利的工具。其目标是快速绘制蛋白质之间的蛋白质相互作用图谱，这可能在控制神经干细胞的增殖和分化中发挥作用。其目标是生成/识别神经干细胞表面蛋白的所有可能的相互作用伙伴，并确认它们在活细胞中的功能。其方法是利用单噬菌体展示来筛选神经干细胞的 cDNA 文库，以分离相应的相互作用伙伴。单噬菌体展示 (SPD) 独特地结合了微加工、噬菌体展示和单分子检测和表征技术，例如荧光相关光谱 (FCS)。该概念基于 1) 文库解决方案的区室化（通过微通道或微滴）达到每个区室在统计上仅容纳单个噬菌体颗粒的水平； 2) 通过 FCS 询问每个区室中噬菌体颗粒的结合状态，以及 3) 通过噬菌体展示鉴定与靶标结合的噬菌体颗粒。为了确保“按需”识别相互作用伙伴的能力，还提出了不带 FCS 的单噬菌体展示作为备用方法。