The goal of this research program is develop a sensitive assay of gene expression patterns which can be used on a variety of scales to study aspects of gene expression in disease, development, physiology and pharmacology. The approach, known as RAGE (Restriction Analysis of Gene Expression), is based on degenerate primer PCR of gene families followed by a restriction endonuclease analysis which allows rapid identification and quantitation of numerous individual genes simultaneously. The RAGE technique is complementary to other current gene expression methods, such as microarray hybridization, SAGE, differential display and RDA; sharing some of the strengths of each of these approaches. Alterations in gene expression due to genomic deletions, translocations, amplifications or drug/hormone treatments, or changes in the transcriptional state of small samples of cells can be easily detected using this approach. The project will expand the range of this technique to numerous gene families involved in signal transduction and cell regulation, such as protein tyrosine kinases, serine/threonine kinases, tyrosine phosphatases, proteases, GTPases, receptor families, and transcription factor families. Additionally, fluorescence based technologies will be applied to increase both the number of gene families and the number of samples which can be easily analyzed. Along with the development of the technology, the project will provide two useful types of datasets of the research community. The first are datasets of restriction fragment lengths for the various gene families necessary to interpret experimental results. The second are datasets of gene expression patterns measured by RAGE on a set of reference samples: the NCI 60 human cancer cell lines being characterize for drug susceptibility. These profiles of gene expression could provide valuable diagnostic and prognostic markers for many types of common human cancers as well as useful information for drug development and evaluation.
|Effective start/end date||9/30/98 → 7/31/03|
- National Institutes of Health: $168,591.00
- National Institutes of Health: $173,016.00
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