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.

抽象的氧化还原生物学越来越多地被认为是细胞信号传导中的关键主题之一。专业该领域的障碍是缺乏监视和测量现场明确定义的氧化还原过程的功能细胞（包括活细胞中的亚细胞结构域），组织或组织。这个4年项目的目标，响应于17-045年的“重点技术研发”的提交是为了扩展红色和远红色荧光生物传感器的荧光蛋白工具箱，可用于成像活细胞和生物中的分室基于硫醇的氧化还原信号传导。在这个项目中，我们旨在在以下方面推进荧光生物传感器技术：（1）我们将扩展基于荧光蛋白的生物传感器，以进行其他氧化还原参数，包括硫氧还蛋白和蛋白质二硫化物异构酶（PDI）在各种细胞室中，例如核，细胞质，，线粒体和内质网（ER）；（2）我们将扩展基于荧光蛋白的颜色红色和远红色的氧化还原生物传感器；（3）我们将验证这些新的生物传感器，并证明使用基于荧光 - 蛋白质的生物传感器，具有兼容颜色，用于多域/多重成像。因为不需要诱导细胞内氧化的紫外线或蓝光来激发红色和远处红色荧光生物传感器，我们的工作将增加基于荧光蛋白的氧化还原的生物相容性生物传感器。此外，与之相比绿色，它们减少了滥用组织和散射。该项目将导致重要的成像工具，以加速对氧化还原的理解调节和信号传导。与基于荧光蛋白的激酶传感器的变革性影响一样，我们预计这些氧化还原生物传感器将启用并改变大量的基本和翻译研究。