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有利的工具。其目标是快速绘制蛋白质之间的相互作用，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/识别神经干细胞表面蛋白的所有可能的相互作用伙伴，并确认其在活细胞中的功能。其方法是利用单噬菌体展示技术筛选神经干细胞cDNA文库，分离相应的相互作用伙伴。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））的独特结合。这个概念是基于1)库溶液的区隔化（通过微通道或微液滴），使每个区隔在统计上仅容纳单个噬菌体颗粒；2)通过FCS对噬菌体颗粒在每个单独的室中的结合状态进行询问，3)通过噬菌体展示识别与靶标结合的噬菌体颗粒。为了确保“按需”识别交互伙伴的能力，还提出了不带FCS的单菌体显示作为备用方法。

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.