Nanodots for Real-Time Monitoring of Signaling Pathways

描述（由申请人提供）：在活细胞中真实的时间监测信号通路的能力将提供用于鉴定和评估信号转导级联调节剂的新的实验方法。本提案的目的是生产、功能化和验证新型蛋白质和肽缀合的纳米颗粒报告物的效用，所述报告物将容易地穿过质膜，在细胞质中浓缩并响应于信号转导事件。将通过光学跟踪荧光信号转导结构域缀合的纳米颗粒至特定的信号依赖性细胞内位置的易位来实时检测信号传导事件。在第二阶段，我们将扩大试验，允许多重分析的细胞信号蛋白的调节，在实时将提供有关信号级联的相互作用的见解，并提供一个独特的生物传感器的来源。这些生物传感器将直接用于评估暴露于潜在的化学或生物威胁剂，识别有毒物质，并发现细胞信号通路的新修饰剂。 在第一阶段提案中开发的工具将帮助医学研究人员了解复杂的生物系统，从而为开发新的诊断或治疗化合物提供机会。

DESCRIPTION (provided by applicant): The ability to monitor signaling pathways in live cells in real time will provide a new experimental methodology for identifying and assessing modulators of signal transduction cascades. The objective of this proposal is to produce, functionalize, and verify the utility of a new type of protein and peptide conjugated nanoparticle reporter that will readily traverse the plasma membrane, concentrate in the cytoplasm and be responsive to signal transduction events. Signaling events will be detected in real-time by optically tracking the translocation of the fluorescent signal transduction domain conjugated nanoparticles to specific, signal dependent intracellular locations. In Phase II we will expand the assay to allow multiplex analysis of the regulation of cell signaling proteins in real-time will provide insight regarding the interplay of signaling cascades and provide a source for unique biosensors. These biosensors will be immediately relevant for assessing exposure to potential chemical or biological threat agents, identifying toxic substances, and discovering novel modifiers of cell signaling pathways. The tools developed in this Phase I proposal will aid medical researchers in their understanding of complex biological systems, thus providing opportunities for the development of new diagnostic or therapeutic compounds.