1. Activity profile-based siRNA screen to explore the functional genomics of Alzheimer's disease
- Author
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Xiao-Ping Shi, Ken S. Koblan, Thomas W. Rosahl, Adam J. Simon, Dirk Beher, Berta Strulovici, Min Xu, Eric Minch, Ira Hoffman, Shane Marine, Daria J. Hazuda, Mark S. Shearman, Stacey Szymanski, John Majercak, David J. Stone, Marc Ferrer, Ansu Bagchi, Amy S. Espeseth, Peter Chase, William J. Ray, Adam Gates, Erica Stec, and Steven R. Bartz
- Subjects
Small interfering RNA ,RNA interference ,HEK 293 cells ,Biological activity ,Human genome ,Transfection ,Biology ,Functional genomics ,Gene ,Molecular biology ,General Biochemistry, Genetics and Molecular Biology ,Biotechnology ,Cell biology - Abstract
Multiparametric assays generate biological activity profiles that provide valuable insight into complex disease models. The use of multiple assay measurements in RNA interference (RNAi) high-throughput screening (HTS) provides biological signatures produced by knocking down individual genes. This strategy has been applied to a genome-wide high-throughput small interfering RNA (siRNA) screen measuring proteolysis of β-amyloid precursor protein (APP) into amyloid β peptides, a critical step in the pathogenesis of Alzheimer’s disease. The assay measures amounts of secreted Aβ40, Aβ42, sAPPα, and sAPPβ from HEK 293 cells stably expressing an optimized APP construct following siRNA transfection. The effect of each siRNA on the four different APP products was simultaneously measured in order to identify human genes that regulate the amyloidogenic processing of APP. Genes with BACE-like and γ-secretase-like activity profiles were identified for further biological characterization.
- Published
- 2007
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