SBIR Phase I: Single Cell Reporter Gene Assay for Hormone Receptors

这个小企业创新研究第一阶段项目旨在通过结合凝胶微滴封装技术和荧光激活细胞分选以及分泌酶捕获和检测格式，开发一种敏感、高通量的报告基因检测方法。报告基因检测用于研究高等生物发育所必需的真核基因表达。尽管报告基因检测已经取得了重大进展，但仍然需要一种高灵敏度、高通量和单细胞分辨率的检测方法。通常，由于丰度低或不稳定，基因产物可能难以检测。此外，许多调节转录的潜在治疗分子表现出较弱的靶标结合亲和力，导致报告基因的低表达。所提出的分析将允许从单个转染细胞中检测广泛的信号强度，这些细胞可以被分离出来用于随后的克隆。I期研究将使用一个模型系统来识别核激素受体的激动剂，核激素受体是控制高等真核生物多种细胞过程的调节开关。提议的分析将通过允许快速筛选针对核激素受体的组合化学或天然产物文库来促进药物发现。所提出的分析将提供一种快速、高通量的方法来筛选潜在治疗分子的大化合物文库。治疗学在全球估计有1000亿美元的市场。

This Small Business Innovation Research Phase I project aims to develop a sensitive, high throughput reporter gene assay by combining gel microdrop encapsulation technology and fluorescence activated cell sorting with a secreted enzyme capture and detection format. Reporter gene assays are used to study eukaryotic gene expression essential for the development of higher organisms. Although there have been significant developments in reporter gene assays, an assay which provides high sensitivity, high throughput and single cell resolution is still needed. Frequently, gene products may be difficult to detect due to low abundance or instability. In addition, many potential therapeutic molecules that modulate transcription, exhibit weak target binding affinities, resulting in low reporter gene expression. The proposed assay will permit detection of a wide range of signal intensities from single, transfected cells which can be isolated for subsequent cloning. Phase I studies will use a model system to identify agonists to nuclear hormone receptors, which are regulatory switches that control diverse cellular processes in higher eukaryotes. The proposed assay will facilitate drug discovery by permitting rapid screening of combinatorial chemistry or natural product libraries against nuclear hormone receptors.The proposed assay will provide a rapid, high throughput method for screening large libraries of compounds for potential therapeutic molecules. Therapeutics is estimated to be a $100 billion market worldwide.