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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：
8550032
负责人：
MATTHEW LEVY
金额：
$ 20.25万
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 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8550032
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.

描述（由申请人提供）：分泌蛋白通常是疾病相关细胞状态（包括ER应激和转移）变化的稳健标志物。缺乏用于检测活细胞中的特定管腔分泌蛋白的技术代表了一个主要差距在细胞成像工具中。我们的目标是开发和提供高灵敏度的报告，可以检测在活细胞群体内的诊断分泌蛋白的表达差异。为了做到这一点，我们将联合收割机结合现有的三种技术，创造一种新的成像工具，STAB（分泌靶向适体BETAB）。更具体地说，通过将修饰的细菌毒素与对分泌蛋白如VEGF或UPR诱导的内质网蛋白Ero 1和ERdj 4具有特异性的核酸酶稳定的适体信标组合，我们的目标是产生试剂，当将其直接加入细胞或组织时，将进入分泌途径并报告这些蛋白的存在。理论上，多达四种不同的荧光染料可以与独特的适体配对，以报告四种不同分泌蛋白的表达。我们设想探针技术将用于组织样本的基础研究和快速临床分析。