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In Vitro Transcription of Long RNA Containing Modified Nucleosides

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Book cover Synthetic Messenger RNA and Cell Metabolism Modulation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 969))

Abstract

The in vitro synthesis of long RNA can be accomplished using phage RNA polymerase and template DNA. However, the in vitro synthesized RNA, unlike those transcribed in vivo in cells, lacks nucleoside modifications. Introducing modified nucleosides into in vitro transcripts is important because they reduce the potential of RNA to activate RNA sensors [1–6] and translation of such nucleoside-modified RNA is increased in cell lines, primary cells, and after in vivo delivery [1, 3, 7–10]. Here, we describe the in vitro synthesis of nucleoside-modified RNA with enhanced translational capacity and reduced ability to activate immune sensors.

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Acknowledgments

These studies were funded by the National Institutes of Health (grant numbers HL87688 to K.K. and AI050484, AI090788, and DE019059 to D.W).

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Authors and Affiliations

  1. Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Norbert Pardi & Drew Weissman

  2. Department of Neurosurgery, University of Pennsylvania, Philadelphia, PA, USA

    Hiromi Muramatsu & Katalin Karikó

Authors
  1. Norbert Pardi
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  2. Hiromi Muramatsu
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  3. Drew Weissman
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  4. Katalin Karikó
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Corresponding author

Correspondence to Katalin Karikó .

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Editors and Affiliations

  1. The Anlyan Center, Department of Genetics, Yale School of Medicine, Cedar Street 300, New Haven, 06520-8005, Connecticut, USA

    Peter M. Rabinovich

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Pardi, N., Muramatsu, H., Weissman, D., Karikó, K. (2013). In Vitro Transcription of Long RNA Containing Modified Nucleosides. In: Rabinovich, P. (eds) Synthetic Messenger RNA and Cell Metabolism Modulation. Methods in Molecular Biology, vol 969. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-260-5_2

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  • DOI: https://doi.org/10.1007/978-1-62703-260-5_2

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-259-9

  • Online ISBN: 978-1-62703-260-5

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