06/19/98 A major application of microarray technology is quantitative monitoring of gene expression patterns. Microarray experiments have been described recently in which dozens of new differentially expressed genes have been discovered using microdissected tissues and radioactive labeling and detection methods. Fluorescent detection methods afford the bast quantitation, permit co-hybridization of two samples labeled with different fluors to the same microarray grid, and permits resolution of very closely spaced microarrayed spots. However, it does have the drawback of increased expense and requires a much higher amount of starting material than radioactive detection methods. This proposal will support the purchase of instrumentation for a state-of-the-art facility that will use microarray technology for genomic analyses. Two instruments will be purchased: a confocal laser-scanning and imaging devise for reading high-density microarrays hybridized with probes labeled with fluorescent tags and a phosphoimager for high- resolution scanning of microarrays hybridized with probes labeled with radioisotopes. The research to be supported by this technology include combining microarray analysis with engineered transcriptional repressors for identification of target genes, fundamental studies of transcription regulation in yeast, studies of cellular pathways involved in transformation and apoptosis, novel approaches to elucidate the biological function of subnuclear cytological structures, studies of tissue regeneration and T-cell development in mice, analysis of subtelomeric variation and transcription patterns in humans, and the development of novel peptide-nucleic acid detection strategies. Graduate students and postdoctoral associates at the Wistar Institute will have access to this emerging technology, allowing them to incorporate this powerful new methodology into their research projects as they develop. More importantly, they will see the exciting new opportunities microarray analysis offers for understanding basic biological processes, and will begin to develop fresh approaches to biological problems that may have been seen as difficult or insoluble using non-global approaches.
