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High-Resolution Gene Expression Profiling of RNA Synthesis, Processing, and Decay by Metabolic Labeling of Newly Transcribed RNA Using 4-Thiouridine

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Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1507))

Abstract

Cellular RNA levels are orchestrated by highly regulated processes involving RNA synthesis (transcription), processing (e.g., splicing, polyadenylation, transport), and degradation. Profiling these changes provides valuable information on the regulation of gene expression. Total cellular RNA is a poor template for revealing short-term changes in gene expression, alterations in RNA decay rates, and the kinetics of RNA processing as well as the differentiation thereof. Here, we describe the metabolic labeling and purification of newly transcribed RNA with 4-thiouridine, by which these limitations are overcome.

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Acknowledgment

This work was supported by MRC fellowship grant G1002523 and NHSBT grant WP11-05 to L.D.

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Authors and Affiliations

  1. Department of Medicine, University of Cambridge, Box 157, Level 5, Addenbrooke’s Hospital, Hills Road, CB20QQ, Cambridge, UK

    Andrzej J. Rutkowski & Lars Dölken

  2. Institut für Virologie und Immunbiologie, Julius-Maximilians-Universität Würzburg, Versbacher Str. 7, 97078, Würzburg, Germany

    Lars Dölken

Authors
  1. Andrzej J. Rutkowski
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  2. Lars Dölken
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Corresponding author

Correspondence to Lars Dölken .

Editor information

Editors and Affiliations

  1. Department of Pathology, Yale University School of Medicine, New Haven, Connecticut, USA

    Narendra Wajapeyee

  2. Department of Pathology, Yale University School of Medicine, New Haven, Connecticut, USA

    Romi Gupta

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Rutkowski, A.J., Dölken, L. (2017). High-Resolution Gene Expression Profiling of RNA Synthesis, Processing, and Decay by Metabolic Labeling of Newly Transcribed RNA Using 4-Thiouridine. In: Wajapeyee, N., Gupta, R. (eds) Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation. Methods in Molecular Biology, vol 1507. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6518-2_10

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  • DOI: https://doi.org/10.1007/978-1-4939-6518-2_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6516-8

  • Online ISBN: 978-1-4939-6518-2

  • eBook Packages: Springer Protocols

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