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High-Throughput siRNA Screening Applied to the Ubiquitin–Proteasome System

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Proteostasis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1449))

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.

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Acknowledgmevnts

The authors apologize to those whose work was not cited due to space constraints. S.V.N., E.G.P., and R.H.P. are supported financially by grants from the Lundbeck Foundation and the Danish Council for Independent Research (Natural Sciences). The authors would like to acknowledge networking support by the Proteostasis COST Action (BM1307).

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    Authors and Affiliations

    1. Department of Biology, Section for Biomolecular Sciences, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen N, 2200, Denmark

      Esben G. Poulsen, Sofie V. Nielsen & Rasmus Hartmann-Petersen Ph.D.

    2. Biotech Research and Innovation Centre, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen N, 2200, Denmark

      Elin J. Pietras, Jens V. Johansen & Cornelia Steinhauer

    Authors
    1. Esben G. Poulsen
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    2. Sofie V. Nielsen
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    3. Elin J. Pietras
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    4. Jens V. Johansen
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    5. Cornelia Steinhauer
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    6. Rasmus Hartmann-Petersen Ph.D.
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    Corresponding author

    Correspondence to Rasmus Hartmann-Petersen Ph.D. .

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    Editors and Affiliations

    1. Computational and Experimental Biology Group Department of Health Promotion and Chronic Diseases, National Health Institute Dr. Ricardo Jorge, INSA, I.P., Lisboa, Portugal

      Rune Matthiesen

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    Poulsen, E.G., Nielsen, S.V., Pietras, E.J., Johansen, J.V., Steinhauer, C., Hartmann-Petersen, R. (2016). High-Throughput siRNA Screening Applied to the Ubiquitin–Proteasome System. In: Matthiesen, R. (eds) Proteostasis. Methods in Molecular Biology, vol 1449. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3756-1_28

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    • DOI: https://doi.org/10.1007/978-1-4939-3756-1_28

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    • Publisher Name: Humana Press, New York, NY

    • Print ISBN: 978-1-4939-3754-7

    • Online ISBN: 978-1-4939-3756-1

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