Optical probes of cardiac channel function

描述(由申请人提供)：离子通道是许多人类病理生理疾病的核心。离子通道异常是人类疾病的主要原因，无论是由于离子通道疾病、药物的副作用，还是由于组织的病理重塑。癫痫、偏头痛、心律失常、囊性纤维化和一些兴奋性疾病是人类的主要疾病。因此，潜在的渠道异常是密集药物开发的重点。然而，目前还没有系统的高通量(甚至中等通量)的方法来筛选离子通道活性的化合物文库。已经开发了许多改变离子通道功能的药物，大多数都是通过基础科学研究或偶然发现的。该项目旨在开发一种能够产生离子通道功能的荧光细胞分析的平台技术。这些检测方法将用于化合物文库的高通量筛选。通过在目标通道的序列中随机插入荧光蛋白序列并随后分析这些插入文库以获得报告活性，从而开发出报告程序。这个第一阶段的项目将通过开发一个人类乙醚-去-去-相关基因(HERG)通道构造库来建立概念验证，并测试这些通道的功能。第二阶段将开发商业技术，允许大规模和高通量开发和筛选插入文库，以开发对任何数量的病理生理重要离子通道的探针

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