A HTS GENE EXPRESSION ASSAY FOR SCREENING NUTRACEUTICALS

The goal of this project is to create a high throughput, sensitive, quantitative and cost-effective platform for measuring gene expression in response to natural compounds derived from foods -nutraceuticals. This technology, the RNA InvaderTM assay, offers highly sensitive detection of mRNA messages directly from cell lysates without the need for reverse transcription or target amplification. We seek to demonstrate the feasibility of using the RNA Invader assay to quantitate the expression of apoptosis-related genes in a variety of cancer cell lines exposed to various nutraceutical compounds through three specific aims: (1) to develop biplex RNA Invader assays that simultaneously quantitate expression of an apoptotic gene and an internal housekeeping gene (2) to evaluate the performance of the assays on an automated platform and determine whether gene expression levels can be quantitated in lysate samples from various cell lines and (3) validate the ability of the Invader RNA assay to measure the changes in gene expression levels in cells exposed to nutraceutical compounds. In Phase 1, we will adapt the format to a broader range of cell lines, candidate genes and nutraceutical compounds.