High-throughput siRNA screening applied to the ubiquitin-proteasome system

Esben Guldahl Poulsen, Sofie V. Nielsen, Elin J. Pietras, Jens Vilstrup Johansen, Cornelia Steinhauer, Rasmus Hartmann-Petersen

2 Citations (Scopus)

Abstract

The ubiquitin-proteasome system is the major pathway for intracellular protein degradation in eukaryotic cells. Due to the large number of genes dedicated to the ubiquitin-proteasome system, mapping degradation pathways for short lived proteins is a daunting task, in particular in mammalian cells that are not genetically tractable as, for instance, a yeast model system. Here, we describe a method relying on high-throughput cellular imaging of cells transfected with a targeted siRNA library to screen for components involved in degradation of a protein of interest. This method is a rapid and cost-effective tool which is also highly applicable for other studies on gene function.

Original languageEnglish
Title of host publicationProteostasis
EditorsRune Matthiesen
Number of pages19
PublisherSpringer
Publication date1 Sept 2016
Pages421-439
Chapter28
ISBN (Print)978-1-4939-3754-7
ISBN (Electronic)978-1-4939-3756-1
DOIs
Publication statusPublished - 1 Sept 2016
SeriesMethods in Molecular Biology
Volume1449
ISSN1064-3745

Keywords

  • Journal Article

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