High-content RNA interference assay: Analysis of tau hyperphosphorylation as a generic paradigm

Rilee H. Robeson, Travis Dunckley

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationDisease Gene Identification
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages221-238
Number of pages18
ISBN (Print)9781617379536
DOIs
StatePublished - Jan 1 2011
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume700
ISSN (Print)1064-3745

Keywords

  • Alzheimer's disease
  • High-content assay
  • Immunofluorescence
  • RNA interference
  • Tau
  • siRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Robeson, R. H., & Dunckley, T. (2011). High-content RNA interference assay: Analysis of tau hyperphosphorylation as a generic paradigm. In Disease Gene Identification: Methods and Protocols (pp. 221-238). (Methods in Molecular Biology; Vol. 700). Humana Press Inc.. https://doi.org/10.1007/978-1-61737-954-3_15