High-Throughput siRNA Screening Applied to the Ubiquitin-Proteasome System.
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.
Department of Biology, Section for Biomolecular Sciences, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen N, 2200, Denmark.
Esben G
EG
Poulsen
Department of Biology, Section for Biomolecular Sciences, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen N, 2200, Denmark.
Sofie V
SV
Nielsen
Biotech Research and Innovation Centre, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen N, 2200, Denmark.
Elin J
EJ
Pietras
Biotech Research and Innovation Centre, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen N, 2200, Denmark.
Jens V
JV
Johansen
Biotech Research and Innovation Centre, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen N, 2200, Denmark.
Cornelia
C
Steinhauer
Department of Biology, Section for Biomolecular Sciences, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen N, 2200, Denmark. rhpetersen@bio.ku.dk.
Rasmus
R
Hartmann-Petersen
D000818:Animals; D006801:Humans; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism*; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism*
D034741:RNA, Small Interfering; D025801:Ubiquitin; D046988:Proteasome Endopeptidase Complex
Degradation; Proteasome; Screening; Ubiquitin; siRNA