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PLENARY PRESENTATION
MONDAY 18 NOVEMBER
15:00 – 15:30 hrs
GENE TRAP MUTAGENESIS FOR THE ANALYSIS OF SIGNALING PATHWAYS
P Soriano
Fred Hutchison
Cancer Research Center
Co-Authors: Chen
W
Institutions:
Fred Hutchison Cancer Research Center
Draft sequences of human and mouse genomes are now available in public databases awaiting functional annotation. Gene trap mutagenesis provides three types of information not readily attainable by other random mutagenesis schemes: mutant phenotype, trapped sequence, and reporter expression. However, the reliability, efficiency and spectrum of current expression-driven screens are restrained by the reliance on reporter gene expression in ES cells. To circumvent this limitation, we have designed a new expression-driven screening platform by combining gene trapping with microarray technology . With a specifically-designed gene trap and a high throughput 3' RACE procedure, 3' cDNA flanks of trapped genes are cloned to construct a spotted cDNA array. The gene trap array thus permits monitoring of the endogenous expression of trapped genes rather than the expression of tagged reporters in any cell type, and the screening scope is far extended with the broad applications of microarrays. This technology is being used to identify and mutate physiologically important targets of the PDGF signaling pathway.
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