Practical High Througput Low Cost Gene Expression Analysis

DESCRIPTION (provided by applicant): Proposed here is the development and application of a high-throughput, multiplexed PCR-based approach to gene expression analysis to broadly enable the use of gene expression profiling for drug research, development and high content screening. The proposed project builds off of the use of two key technologies, Althea's multiplexed XP PCR, and Blueshift's IsoCyte high performance plate scanner. These two technologies are combined in the proposed feasibility study through the use of a novel approach to preparing a barcoded random bead array for monitoring the gene expression endpoints. The end goal is a method for the performance of multiplexed gene expression analysis of 20-40 genes per sample with a scanning throughput of 1536 samples in 2 minutes. The pace at which the genetics of complex diseases, such as cancer, diabetes, cardiovascular and inflammatory disorders, are being deciphered has been accelerating. Researchers have begun to characterize in detail multiple genetic mechanisms that give rise to these complex diseases, as well as numerous functional pathways associated with disease response such as stress and damage response, immune response, cell cycle regulation, cell proliferation and cell death. This exponential growth in our knowledgebase of disease genetics has led to the identification of a large array of genes, proteins and pathways that potentially play a central role in disease development and may be potential targets for therapeutic intervention. The challenge now is to experimentally delve deeper, both into how these genes may function pathologically and protectively in disease, and into how different compounds and compound classes might influence these genes.