NSF Award
9960154
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.<br/><br/>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.
