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Red and Far-Red Fluorescent Biosensors for Compartmentalized Redox Signaling

用于分隔氧化还原信号传导的红色和远红荧光生物传感器

基本信息

批准号：
10222722
负责人：
Huiwang Ai
金额：
$ 37.84万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-05 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10222722
关键词：
Aging Animal Model Basic Science Biology Biosensor Cancer Etiology Cell Compartmentation Cell Nucleus Cell model Cells Chemicals Color Coupled Coupling Cytosol Detection Development Disease Doxorubicin Drug Screening Endoplasmic Reticulum Engineering Epidermal Growth Factor FDA approved Fluorescence Glutathione Goals Human Hydrogen Peroxide Image Imaging Device Individual Inflammatory Intervention Laboratories Lead Libraries Light Measurement Measures Methods Mitochondria Molecular Monitor Nitrogen Nuclear Oncology Organism Oxidation-Reduction Oxidative Stress Oxygen Penetration Pharmaceutical Preparations Phosphotransferases Photons Physiological Process Protein Disulfide Isomerase Proteins Regulation Reporting Research Sensory Signal Transduction Stimulus Sulfhydryl Compounds Sulfur TXN gene Technology Tissues Toxicology Work absorption animal imaging base biomaterial compatibility chemotherapy multiplexed imaging mutant novel novel therapeutics oxidation public health relevance ratiometric red fluorescent protein response sensor spatiotemporal subcellular targeting technology research and development tool toxicant translational study whole animal imaging

项目摘要

ABSTRACT Redox biology has been increasingly recognized as one of the key themes in cell signaling. A major obstacle in this field is the lack of capabilities to monitor and measure clearly defined redox processes in live cells (including subcellular domains in live cells), tissues, or organisms. The goal of this 4-year project, submitted in response to PAR-17-045 “Focused Technology Research and Development”, is to expand the fluorescent protein toolbox for red and far-red fluorescent biosensors that can be used to image compartmentalized, thiol-based redox signaling in live cells and organisms. In this project, we aim to advance the fluorescent biosensor technology in the following aspects : (1) we will expand fluorescent-protein-based biosensors for additional redox parameters, including the redox of thioredoxin and protein disulfide isomerase (PDI) in various cell compartments, such as the nucleus, cytosol, mitochondria, and endoplasmic reticulum (ER); (2) we will expand the colors of fluorescent-protein-based redox biosensors to red and far-red; (3) we will validate these new biosensors and demonstrate the use of fluorescent-protein-based biosensors in compatible colors for multicompartment/multiplex imaging. Because UV or blue light, which can induce intracellular oxidation, is not needed to excite red and far- red fluorescent biosensors, our work will increase the biocompatibility of fluorescent-protein-based redox biosensors. Furthermore, red and far-red biosensors will assist in whole-animal imaging, since compared to green ones, they reduce tissue absorption and scattering. This project will lead to important imaging tools that may accelerate the understanding of redox regulation and signaling. As with the transformative impact of fluorescent-protein-based kinase sensors, we expect that these redox biosensors will enable and transform a large array of basic and translational studies.

摘要氧化还原生物学已经越来越被认为是细胞信号传导的关键主题之一。一个主要该领域中的障碍是缺乏监测和测量活体中明确定义的氧化还原过程的能力细胞（包括活细胞中的亚细胞结构域）、组织或生物体。这个为期四年的项目的目标，提交的响应PAR-17-045“重点技术研究和开发”，是为了扩大用于红色和远红色荧光生物传感器的荧光蛋白工具箱，可用于成像在活细胞和生物体中的区室化的、基于硫醇的氧化还原信号传导。在本项目中，我们的目标是在以下方面推进荧光生物传感器技术：（1）我们将扩大荧光蛋白为基础的生物传感器额外的氧化还原参数，包括氧化还原硫氧还蛋白和蛋白质二硫键异构酶（PDI）在各种细胞区室，如细胞核，胞质溶胶，线粒体、内质网（ER）;（2）我们将扩大荧光蛋白的颜色基础氧化还原生物传感器的红色和远红;（3）我们将验证这些新的生物传感器，并证明使用基于荧光蛋白的生物传感器，颜色兼容，用于多室/多重成像。因为可以诱导细胞内氧化的紫外光或蓝光不需要激发红光和远红光，红色荧光生物传感器，我们的工作将增加生物相容性的荧光蛋白基氧化还原生物传感器此外，红色和远红色生物传感器将有助于整个动物成像，因为与绿色的，它们减少组织吸收和散射。这个项目将导致重要的成像工具，可能会加速氧化还原的理解调节和信号。与基于荧光蛋白的激酶传感器的变革性影响一样，预计这些氧化还原生物传感器将使大量的基础和转化研究成为可能，并改变这些研究。