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HPLC Purification of In Vitro Transcribed Long RNA

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Synthetic Messenger RNA and Cell Metabolism Modulation

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

Abstract

In vitro transcription of DNA with phage RNA polymerases is currently the most efficient method to produce long sequence-specific RNA. While the reaction can yield large quantities of RNA, it contains impurities due to various unwanted activities of the polymerases. Here, we described an easily performed HPLC purification that removes multiple contaminants from in vitro transcribed RNA and is scalable. The purified RNA is translated at much greater levels, especially in primary cells and in vivo. HPLC purification of RNA containing modified nucleosides that suppress RNA-mediated activation of innate immune sensors leads to a non-immunogenic RNA with superior translational capacity.

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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

    Drew Weissman & Norbert Pardi

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

    Hiro Muramatsu & Katalin Karikó

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

Correspondence to Drew Weissman .

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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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Weissman, D., Pardi, N., Muramatsu, H., Karikó, K. (2013). HPLC Purification of In Vitro Transcribed Long RNA. 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_3

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  • DOI: https://doi.org/10.1007/978-1-62703-260-5_3

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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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