Optical probes of cardiac channel function

心经功能光学探针

• 批准号: 6742915
• 负责人: Vincent A Pieribone
• 金额: $ 9.95万
• 依托单位: PHYSIOLOGICAL TECHNOLOGIES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-15 至 2006-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6742915
• 关键词: chemical registry /resource; fluorescent dye /probe; high throughput technology; ion channel blocker; membrane activity; membrane channels; molecular probes; technology /technique development; transfection; transposon /insertion element

DESCRIPTION (provided by applicant): Ion channels are at the heart of a number of human pathophysiological conditions. Either as channelopathies, side effects of drugs, or as a result of pathological remodeling of tissues, ion channel irregularities account for a major segment of human disease. Epilepsy, migraine, cardiac arrhythmias, cystic fibrosis, and a number of excitability diseases represent major human maladies. As such the underlying channel abnormalities are the focus of intensive drug development. However, currently there is no systematic high-throughput (or even medium throughput) method of screen compound libraries for ion channel activity. There are a number of pharmaceuticals that have been developed that alter ion channel function most have be discovered by basic scientific studies or serendipitously. This project seeks to develop a platform technology that can generate fluorescent cellular assays of ion channel function. These assays will be used in high-throughput screens of compound libraries. The reporters are developed by the random insertion of a fluorescent protein sequence within the target channel's sequence and the subsequent analysis of these insertion libraries for reporter activity. This Phase I project will establish the proof of concept by developing a library of human ether-a-go-go related gene (HERG) channel constructs and test these channels for function. Phase II will develop the commercial technology that will allow the large scale and high throughput development and screening of insertional libraries to develop probes to any number of pathophysiologically important ion channels

描述（由申请人提供）：离子通道是许多人类病理生理状况的核心。无论是作为通道病、药物的副作用，还是作为组织的病理性重塑的结果，离子通道不规则性都是人类疾病的主要部分。癫痫、偏头痛、心律失常、囊性纤维化和许多兴奋性疾病代表了人类的主要疾病。因此，潜在的通道异常是密集药物开发的焦点。然而，目前还没有系统的高通量（或甚至中等通量）筛选化合物文库的离子通道活性的方法。已经开发出许多改变离子通道功能的药物，大多数是通过基础科学研究或偶然发现的。该项目旨在开发一种平台技术，可以生成离子通道功能的荧光细胞测定。这些测定将用于化合物文库的高通量筛选。通过在靶通道序列内随机插入荧光蛋白序列并随后分析这些插入文库的报告活性来开发报告基因。这个第一阶段项目将通过开发人类ether-a-go-go相关基因（HERG）通道构建体库来建立概念验证，并测试这些通道的功能。第二阶段将开发商业技术，该技术将允许大规模和高通量开发和筛选插入文库，以开发针对任何数量的病理生理学重要离子通道的探针。