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Preparation of Functional, Fluorescently Labeled mRNA Capped with Anthraniloyl-m7GpppG

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Synthetic mRNA

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1428))

Abstract

Fluorescent mRNA molecules offer a wide range of applications for studying capping/decapping reactions, translation, and other biophysical studies. Furthermore, fluorescent tags prove invaluable for tracking RNA molecules in cells. Here, we describe an efficient synthesis of a fluorescent cap analog, anthranioyl-GTP, its purification, and in vitro cap labeling of transcribed mRNA catalyzed by the recombinant vaccinia capping enzyme to produce anthranioyl-m7GpppG-capped RNA.

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Acknowledgment

We thank Jason A. Domashevskiy for his critical review of the manuscript.

Author information

Authors and Affiliations

  1. Department of Sciences, John Jay College of Criminal Justice, City University of New York, New York, NY, USA

    Artem V. Domashevskiy & David J. Rodriguez

  2. Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria, Australia

    Dilantha Gunawardana

  3. Department of Botany, University of Sri Jayewardenepura, Nugegoda, Sri Lanka

    Dilantha Gunawardana

  4. Department of Chemistry, Hunter College, City University of New York, New York, NY, USA

    Dixie J. Goss

Authors
  1. Artem V. Domashevskiy
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  2. David J. Rodriguez
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  3. Dilantha Gunawardana
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  4. Dixie J. Goss
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Corresponding author

Correspondence to Artem V. Domashevskiy .

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Editors and Affiliations

  1. Dept of Biochemistry&Molecular Bio, Louisiana State Univ Health Scien center, Shreveport, Louisiana, USA

    Robert E. Rhoads

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Domashevskiy, A.V., Rodriguez, D.J., Gunawardana, D., Goss, D.J. (2016). Preparation of Functional, Fluorescently Labeled mRNA Capped with Anthraniloyl-m7GpppG. In: Rhoads, R. (eds) Synthetic mRNA. Methods in Molecular Biology, vol 1428. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3625-0_4

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  • DOI: https://doi.org/10.1007/978-1-4939-3625-0_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3623-6

  • Online ISBN: 978-1-4939-3625-0

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