title: High-content RNA interference assay: analysis of tau hyperphosphorylation as a generic paradigm.
abstract: High-content analysis methods provide the opportunity to interrogate specific cellular end points in living cells. When coupled with high-throughput RNA interference (ht-RNAi) loss of function screens, high-content analyses are a powerful discovery tool for the identification of new genes and pathways involved in specific disease-relevant cellular functions. The most common readout is a fluorescence measurement, usually based on a green fluorescent protein reporter (or some derivative thereof ) or a fluorescently labeled antibody. Here, we describe a specific approach to the development of a high-content assay for the hyperphosphorylation of tau protein that is compatible with RNAi screens. The goal of this chapter is to provide a generic paradigm, using hyperphosphorylation of tau protein as an example, to serve as a blueprint for the investigation of additional cellular end points or protein functions for those interested in performing high-content screens.
authors.affiliation: Neurogenomics Division, Translational Genomics Research Institute, Phoenix, AZ, USA.
authors.forename: Rilee H
authors.initials: RH
authors.lastname: Robeson
authors.forename: Travis
authors.initials: T
authors.lastname: Dunckley
mesh_terms: D000544:Alzheimer Disease / Q000235:genetics / Q000503:physiopathology; D045744:Cell Line, Tumor; D005455:Fluorescent Antibody Technique; D055106:Genome-Wide Association Study / Q000379:methods*; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D016875:tau Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D034741:RNA, Small Interfering; D016875:tau Proteins