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：
9975915
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 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9975915
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被认为是细胞信号传导的关键主题之一。该领域的障碍是缺乏监测和测量现场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域）、组织或生物体这个为期 4 年的项目的目标，为响应 PAR-17-045“重点技术研究与开发”而提交，旨在扩大用于可用于成像的红色和远红色荧光生物传感器的荧光蛋白工具箱活细胞和生物体中基于硫醇的区室化氧化还原信号传导。在这个项目中，我们的目标是在以下方面推进荧光生物传感器技术：（1）我们将扩展基于荧光蛋白的生物传感器以获得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）存在于各种细胞区室中，例如细胞核、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光和远红光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演示其用途基于荧光蛋白的生物传感器，具有兼容的颜色，用于多室/多重成像。因为可以诱导细胞内氧化的紫外线或蓝光不需要激发红光和远光红色荧光生物传感器，我们的工作将提高基于荧光蛋白的氧化还原的生物相容性此外，与传统的生物传感器相比，红色和远红色生物传感器将有助于整个动物成像。绿色的，它们减少组织的吸收和散射。该项目将带来重要的成像工具，可以加速对氧化还原的理解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实现并改变大量基础和转化研究。