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Isolation of Newly Transcribed RNA Using the Metabolic Label 4-Thiouridine

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Book cover mRNA Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1648))

Abstract

Isolation of newly transcribed RNA is an invaluable approach that can be used to study the dynamic life of RNA in cellulo. Traditional methods of whole-cell RNA extraction limit subsequent gene expression analyses to the steady-state levels of RNA abundance, which often masks changes in RNA synthesis and processing. This chapter describes a methodology with low cytotoxicity that permits the labeling and isolation of nascent pre-mRNA in cell culture. The resulting isolate is suitable for use in a series of downstream applications aimed at studying changes in RNA synthesis, processing, or stability.

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Acknowledgments

Research in the Hertel laboratory is supported by NIH (GM062287, GM110244 and F31CA17179). Special thanks to Nate Hoverter for contribution of key graphics in Fig. 1.

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Author notes

  1. Angela Garibaldi and Francisco Carranza contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology and Molecular Genetics, University of California, Irvine, B233 Med Sci I, Irvine, CA, 92697-4025, USA

    Angela Garibaldi, Francisco Carranza & Klemens J. Hertel

Authors
  1. Angela Garibaldi
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  2. Francisco Carranza
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  3. Klemens J. Hertel
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Corresponding author

Correspondence to Klemens J. Hertel .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Irvine, California, USA

    Yongsheng Shi

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Garibaldi, A., Carranza, F., Hertel, K.J. (2017). Isolation of Newly Transcribed RNA Using the Metabolic Label 4-Thiouridine. In: Shi, Y. (eds) mRNA Processing. Methods in Molecular Biology, vol 1648. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7204-3_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7204-3_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7203-6

  • Online ISBN: 978-1-4939-7204-3

  • eBook Packages: Springer Protocols

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