International Mammalian Genome Society

The 16th International Mouse Genome Conference (2002)


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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