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m6A RNA Immunoprecipitation Followed by High-Throughput Sequencing to Map N6-Methyladenosine

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Post-Transcriptional Gene Regulation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2404))

Abstract

N6-methyladenosine (m6A) is the most abundant internal modification on messenger RNAs (mRNAs) and long noncoding RNAs (lncRNAs) in eukaryotes. It influences gene expression by regulating RNA processing, nuclear export, mRNA decay, and translation. Hence, m6A controls fundamental cellular processes, and dysregulated deposition of m6A has been acknowledged to play a role in a broad range of human diseases, including cancer. m6A RNA immunoprecipitation followed by high-throughput sequencing (MeRIP-seq or m6A-seq) is a powerful technique to map m6A in a transcriptome-wide level. After immunoprecipitation of fragmented polyadenylated (poly(A)+) rich RNA by using specific anti-m6A antibodies, both the immunoprecipitated RNA fragments together with the input control are subjected to massively parallel sequencing. The generation of such comprehensive methylation profiles of signal enrichment relative to input control is necessary in order to better comprehend the pathogenesis behind aberrant m6A deposition.

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Acknowledgments

This research was supported by grants from the Knut and Alice Wallenberg Foundation, Umeå University, Västerbotten County Council, Swedish Research Council (2017-01636), Kempe Foundation (SMK-1766), and Cancerfonden (19 0337 Pj).

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Authors and Affiliations

  1. Department of Molecular Biology, Umeå University, Umeå, Sweden

    Devi Prasad Bhattarai & Francesca Aguilo

  2. Wallenberg Center for Molecular Medicine, Umeå University, Umeå, Sweden

    Devi Prasad Bhattarai & Francesca Aguilo

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  1. Devi Prasad Bhattarai
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  2. Francesca Aguilo
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Correspondence to Francesca Aguilo .

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Editors and Affiliations

  1. Department of Cellular, Computational and Integrative Biology (CIBIO), University of Trento, Trento, Italy

    Erik Dassi

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Bhattarai, D.P., Aguilo, F. (2022). m6A RNA Immunoprecipitation Followed by High-Throughput Sequencing to Map N6-Methyladenosine. In: Dassi, E. (eds) Post-Transcriptional Gene Regulation. Methods in Molecular Biology, vol 2404. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1851-6_19

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  • DOI: https://doi.org/10.1007/978-1-0716-1851-6_19

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1850-9

  • Online ISBN: 978-1-0716-1851-6

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