Fluorescent Biosensors for Networks and Pathways

项目总结（见说明）： 我们建议继续建立一个全国可见的响应中心，专注于开发新型荧光生物传感器和检测技术，用于在活细胞中真实的时间和高空间分辨率下研究途径和网络。新中心保留了匹兹堡现有两个中心的综合经验和基础设施：卡内基梅隆大学的分子生物传感器和成像中心 匹兹堡大学的生物成像中心。该中心的技术开发（核心1）组件将创建一个强大的细胞内荧光生物传感器和报告工具箱，可用于研究许多（如果不是全部）活细胞中的通路蛋白和活动。这个荧光生物传感器开发计划整合了多个学科的努力，包括染料 化学、分子生物学、生物化学、结构生物学、建模、细胞生物学、图像采集和分析以及高通量筛选。在过去三年的资助期间，我们在这方面取得了重大进展。 四个令人兴奋的驾驶生物学项目（核心2）对核心1的技术开发工作至关重要。 这些DBP专注于重要的和目前无法解决的生物学问题，并将作为该技术的试验台，以及该计划中开发的生物传感器和报告器的价值的引人注目的演示。 基础设施（核心3）由匹兹堡大学生物成像中心提供。CBI的作用是通过测试具有挑战性的生物问题的新生物传感器，作为整个项目的应用和推广机构。在本提案的最后一个周期中，CBI的明确使命演变成探测器实施的催化剂，并加强和扩大MBIC开发的新探测器的影响。这是通过提供设施和 在推动生物项目的背景下测试和验证探针的专业知识，并最终在整个生物医学研究界。 该计划包含一个重要的技术转让组成部分，以传播概念，知识，软件，材料和资源给学术研究实验室和行业的用户。这是通过我们的基础设施、培训和传播核心（核心3、4、5）以及技术转让实现的 管理核心（Core 6）。

PROJECT SUMMARY (See Instructions): We propose to continue a nationally visible and responsive center focused on the development of novel fluorescent biosensor and detection technologies for investigating pathways and networks in real time and high spatial resolution in living cells. The renewed center retains the combined experience and infrastructure of two existing centers in Pittsburgh: the Molecular Biosensors and Imaging Center at Carnegie Mellon University and the Center for Biologic Imaging at the University of Pittsburgh. The Technology Development (Core 1) component of the center will create a powerful toolbox of intracellular fluorescent biosensors and reporters that can be used to study many, if not all, of the pathway proteins and activities in living cells. This fluorescent biosensor development program integrates efforts across multiple disciplines, including dye chemistry, molecular biology, biochemistry, structural biology, modeling, cell biology, image acquisition and analysis, and high throughput screening. We have made substantial progress in this effort during the previous 3 years of funding. Four exciting Driving Biology Projects (Core 2) are essential to the technology development effort in Core 1. These DBPs are focused on important and currently un-addressable biological problems, and will serve both as test-beds for the technology and compelling demonstrations of the value of the biosensors and reporters developed in this program. The Infrastructure (Core 3) is provided through the Center for Biologic Imaging at the University of Pittsburgh. The role of the CBI is to act as the application and outreach ami of the project as a whole by testing the new biosensors with challenging biological problems. During the last cycle of this proposal the clear mission of the CBI evolved to become the catalyst in probe implementation, and to strengthen and broaden the impact of the new probes developed by MBIC. This is achieved by providing facilities and expertise to test and validate the probes in the context of the driving biological projects, and ultimately, the biomedical research community at large. The program contains a significant technology transfer component to disseminate concepts, knowledge, software, materials, and resources to users in both academic research labs and industry. This is achieved in our Infrastructure, Training, and Dissemination cores (Core 3,4, 5) and through the technology transfer activities in the Management core (Core 6).