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Detecting Internal N7-Methylguanosine mRNA Modifications by Differential Enzymatic Digestion Coupled with Mass Spectrometry Analysis

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RNA Modifications

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2298))

Abstract

The recent discovery of reversible chemical modifications on mRNA has opened a new era of post-transcriptional gene regulation in eukaryotes. Among these modifications identified in eukaryotic mRNA, N7-methylguanosine (m7G) is unique owing to its presence in the 5′ cap structure. Recently, it has been reported that m7G also exists internally in mRNA. Here, we describe a protocol of combining differential enzymatic digestion with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis to detect internal m7G modification in mRNA. This protocol can also be used to quantify the level of m7G at both the 5′ cap and internal positions of mRNA.

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Acknowledgments

The work is supported by the National Key R&D Program of China (2017YFC0906800) and the National Natural Science Foundation of China (21672166, 21635006, 21721005).

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Authors and Affiliations

  1. Sauvage Center for Molecular Sciences, Department of Chemistry, Wuhan University, Wuhan, China

    Xue-Jiao You & Bi-Feng Yuan

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  1. Xue-Jiao You
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  2. Bi-Feng Yuan
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Corresponding author

Correspondence to Bi-Feng Yuan .

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Editors and Affiliations

  1. Department of Urology, University of California, San Francisco, San Francisco, CA, USA

    Mary McMahon

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You, XJ., Yuan, BF. (2021). Detecting Internal N7-Methylguanosine mRNA Modifications by Differential Enzymatic Digestion Coupled with Mass Spectrometry Analysis. In: McMahon, M. (eds) RNA Modifications. Methods in Molecular Biology, vol 2298. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1374-0_16

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  • DOI: https://doi.org/10.1007/978-1-0716-1374-0_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1373-3

  • Online ISBN: 978-1-0716-1374-0

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