Nanotechnology for Systems Biology of Neural Stem Cells

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造一个使能工具。它的目标是快速定位蛋白质的蛋白质-蛋白质相互作用，这可能在控制神经干细胞的增殖和分化中发挥作用。其目的是生成/鉴定神经干细胞表面蛋白的所有可能的相互作用伴侣，并确认它们在活细胞中的功能。其方法是利用噬菌体展示技术筛选神经干细胞的cDNA文库，分离相应的相互作用伴侣。单噬菌体展示（SPD）是微加工、噬菌体展示和单分子检测和表征技术如荧光相关光谱（FCS）的独特组合。该概念基于1）文库溶液的区室化（通过微通道或微滴）至每个区室在统计学上仅容纳单个噬菌体颗粒的水平; 2）通过FCS询问每个单独区室中噬菌体颗粒的结合状态;和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”鉴定相互作用伴侣的能力，还提出了不含FCS的单噬菌体展示作为备用方法。

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.