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Journal of Biomolecular Screening, Vol. 9, No. 8, 704-711 (2004)
DOI: 10.1177/1087057104268698

Transcriptional-Based Screens for Pathway-Specific, High-Throughput Target Discovery in Endothelial Cells

Robert L. Yauch

SUGEN, Inc., South San Francisco, CA.,Molecular Diagnostics, Genentech, Inc. Mail Stop 50, 1 DNA Way, South San Francisco, CA 94080.bobyauch{at}gene.com

Edward E. Kadel, III

SUGEN, Inc., South San Francisco, CA.

Cory Nicholas

SUGEN, Inc., South San Francisco, CA.

Selwyna Tetangco

SUGEN, Inc., South San Francisco, CA.

Douglas O. Clary

SUGEN, Inc., South San Francisco, CA.

With the sequence of the human genome at hand, target discovery strategies are needed that can rapidly identify novel gene products involved in human disease pathways. In this article, the authors describe a cell-based, high-throughput assay that can identify gene products capable of modulating the vascular endothelial growth factor (VEGF) and tumor necrosis factor • (TNFa) signaling pathways in human endothelial cells. The assay uses real-time PCRtechnology tomeasure downstreamreporter mRNA transcripts induced upon cytokine stimulation in a 96-well plate format and has been adapted for use with recombinant adenoviruses. The authors specifically demonstratemodulation of cytokine-driven reporter transcripts using drug inhibitors and through adenoviral-mediated expression of known signaling intermediates of the respective pathways. In addition, they have used an arrayed library of 350 recombinant adenoviruses to screen for novel modulators of the VEGF and TNF• pathways. The high-throughput screening capacity and sensitivity of this system make it a useful tool for new drug target identification.

Key Words: functional genomics • adenovirus • VEGF • TNF•


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