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RNAi Screens for Genes Involved in Golgi Glycosylation

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Membrane Trafficking

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1270))

Abstract

RNAi screening has gained popularity in recent years, due to its usefulness in systematic investigations of biological pathways. Combined with high-content screening and advances in imaging and analysis methods, it can enable detailed genetic characterization of cellular processes such as protein glycosylation, a major function of the Golgi apparatus. Glycosylation concerns about one third of all human proteins and regulates various cellular behaviors. Yet the methods available to study it are limited and not easily accessible. In this chapter, we detail a step-by-step method to systematically and quantitatively investigate glycosylation using fluorescent lectin staining, following high-throughput RNAi-based downregulation of gene activities. We also provide a workflow for downstream analysis of the data generated.

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Author information

Authors and Affiliations

  1. Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, 61 Biopolis Drive, Proteos, Singapore, 138673, Singapore

    Germaine Y. Goh & Frederic A. Bard

  2. Department of Biochemistry, National University of Singapore, 21 Lower Kent Ridge Road, Singapore, 119077, Singapore

    Frederic A. Bard

Authors
  1. Germaine Y. Goh
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  2. Frederic A. Bard
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Corresponding author

Correspondence to Frederic A. Bard .

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Editors and Affiliations

  1. Department of Biochemistry, National University of Singapore, Singapore, Singapore

    Bor Luen Tang

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Goh, G.Y., Bard, F.A. (2015). RNAi Screens for Genes Involved in Golgi Glycosylation. In: Tang, B. (eds) Membrane Trafficking. Methods in Molecular Biology, vol 1270. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2309-0_28

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  • DOI: https://doi.org/10.1007/978-1-4939-2309-0_28

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2308-3

  • Online ISBN: 978-1-4939-2309-0

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