Fluorescent probes for quantitation of secretory protein levels in single cells

用于定量单细胞分泌蛋白水平的荧光探针

基本信息

批准号：
8413995
负责人：
MATTHEW LEVY
金额：
$ 25.05万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-24 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8413995
关键词：
Alpha Cell Antibodies Bacterial Toxins Basic Science Beta Cell Binding Cell membrane Cells Chemistry Clinical Complex Cultured Tumor Cells Detection Development Diabetes Mellitus Diagnosis Diagnostic Disease Endoplasmic Reticulum Engineering Environment Enzyme-Linked Immunosorbent Assay Enzymes Fluorescent Antibody Technique Fluorescent Dyes Fluorescent Probes Goals Growth Factor Heart Diseases Imaging Device Immunoblotting Immunofluorescence Immunologic In Vitro Label Lectin Life Link Liver diseases Malignant Neoplasms Mass Spectrum Analysis Membrane Proteins Methods Monitor Mus Neoplasm Metastasis Noise Oligonucleotides One-Step dentin bonding system Pancreas Pathway interactions Population Proteins RNA Reagent Reporter Reporting Secretory Cell Signal Transduction Signaling Molecule Staging Staining method Stains Stress Technology Therapeutic Time Tissue Sample Tissues Vascular Endothelial Growth Factors Work aptamer bevacizumab biological adaptation to stress cell fixing cell type cellular imaging endoplasmic reticulum stress extracellular human disease neoplastic cell nuclease protein expression receptor response sample fixation secretory protein tool

项目摘要

DESCRIPTION (provided by applicant): Secretory proteins are often robust markers of changes in disease-relevant cellular states including ER stress and metastasis. The absence of technologies for detecting specific luminal secretory proteins in live cells represents a major gap in cell imaging tools. Our goal is to develop and deliver highly sensitive reporters that can detec differences in the expression of diagnostic secretory proteins within a population of live cells. T do this, we will combine three existing technologies to create a new class of imaging tools, STABs (Secretory Targeting Aptamer Beacons). More specifically, by combining modified bacterial toxins with nuclease stabilized aptamer beacons specific for secreted proteins such as VEGF or the UPR-induced endoplasmic reticulum proteins Ero1 and ERdj4, we aim to generate reagents which, when added directly to cells or tissues, will enter the secretory pathway and report the presence of these proteins. Theoretically, up to four distinct fluorescent dyes can be paired with unique aptamers to report on the expression of four different secretory proteins. We envision the probe technology will have utility for basic research and rapid clinical analysis of tissue samples. PUBLIC HEALTH RELEVANCE: Secretory proteins are often robust markers of changes in disease-relevant cellular states including ER stress and metastasis. The proposed work seeks to develop new tools to directly detect the presence of or changes in the levels of proteins within the secretory pathway of na¿ve, unperturbed live cells. We envision the probe technology will have utility for basic research and rapid clinical analysis of tissue samples.

描述（由申请人提供）：分泌蛋白通常是疾病相关细胞状态变化的强有力的发光标记，包括内质网应激和转移。缺乏检测活细胞中特定分泌蛋白的技术是一个主要差距。我们的目标是开发和提供能够检测活细胞群中诊断分泌蛋白表达差异的高灵敏度生产器。为此，我们将结合三种现有技术来创建一种新型成像技术。工具，STAB（分泌靶向适体信标）更具体地说，通过将修饰的细菌毒素与特异性针对分泌蛋白（例如 VEGF 或 UPR 诱导的内质）的核酸酶稳定的适体信标相结合。网状蛋白 Ero1 和 ERdj4，我们的目标是生成试剂，当直接添加到细胞或组织中时，这些试剂将进入分泌途径并报告这些蛋白质的存在。理论上，最多四种不同的荧光染料可以与独特的适体配对来报告。我们预计探针技术将可用于组织样本的基础研究和快速临床分析。公共健康相关性：分泌蛋白通常是疾病相关细胞状态变化的有力标志物，包括内质网应激和转移。拟议的工作旨在开发新工具来直接检测细胞内蛋白质水平的存在或变化。 na的分泌途径ve，未受干扰的活细胞，我们预计探针技术将可用于组织样本的基础研究和快速临床分析。