Rapid Analysis of Gene Expression
Current techniques for analysis of gene expression either monitor one gene at a time, for example northern hybridization or RT-PCR methods, or are designed for the simultaneous analysis of thousands of genes, for example microarray hybridization or serial analysis of gene expression. To provide a flexible, intermediate scale alternative, a PCR-based method RAGE (rapid analysis of gene expression) has been developed which allows expression changes to be determined in either a directed search of known genes, or an undirected survey of unknown genes. A single set of reagents and reaction conditions allows analyses of most genes in any eukaryote. The method is useful for assaying on the order of tens to hundreds of genes in multiple samples. Control experiments indicate reliable detection of changes in gene expression 2-fold and greater, and sensitivity of detection better than 1 in 10,000. This technology has been applied to investigate the changes in gene expression in human cells following treatment with a carcinogen and to determine the changes in large numbers of genes in early stage breast cancer. These molecular “signatures” of cancer may then be used to determine which tumors are likely to be responsive to chemotherapy.