FLUORESCENT BIOSENSORS FOR NETWORKS AND PATHWAYS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 描述(由申请人提供)：我们建议继续建立一个全国性的可见和响应中心，专注于开发新型荧光生物传感器和检测技术，以实时和高空间分辨率研究活细胞中的途径和网络。更新后的中心保留了匹兹堡现有的两个中心的合并经验和基础设施：卡内基梅隆大学的分子生物传感器和成像中心和匹兹堡大学的生物成像中心。该中心的技术开发(核心1)部分将创建一个强大的细胞内荧光生物传感器和记者工具箱，可用于研究活细胞中的许多(如果不是全部)途径蛋白质和活动。这一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3年的筹资期间，我们在这方面的努力取得了实质性进展。四个激动人心的驱动生物学项目(核心2)对核心1的技术开发工作至关重要。这些DBP专注于目前无法解决的重要生物问题，将作为技术的试验台，并引人注目地展示在该计划中开发的生物传感器和记者的价值。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。在本提案的最后一个周期中，CBI的明确使命演变成了实施探测的催化剂，并加强和扩大了MBIC开发的新探测的影响。这是通过提供设施和专业知识来在推动生物项目的背景下测试和验证探针，并最终在整个生物医学研究社区中实现的。该计划包含一个重要的技术转让部分，用于向学术研究实验室和行业的用户传播概念、知识、软件、材料和资源。这是在我们的基础设施、培训和传播核心(核心3、4、5)中实现的，并通过管理核心(核心6)中的技术转让活动实现。相关性(见说明)：了解蛋白质在网络和通路中的调节对于诊断、治疗和治愈各种人类疾病至关重要。该计划开发了一套新的工具，用于观察活细胞中这种调控过程的发生，并将导致技术、生物学知识和药物发现的根本性进步。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): 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 acheived 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). RELEVANCE (See instructions): Understanding the regulation of proteins in networks and pathways is central to diagnosing, treating and curing a variety of human diseases. This program develops a new set of tools for observing this regulation process in live cells, as it occurs, and will lead to fundamental advances in technology, biological knowledge, and drug discovery.