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m6A RNA immunoprecipitation followed by high-throughput sequencing to map N6-Methyladenosine
Umeå University, Faculty of Medicine, Wallenberg Centre for Molecular Medicine at Umeå University (WCMM). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).
Umeå University, Faculty of Medicine, Wallenberg Centre for Molecular Medicine at Umeå University (WCMM). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).ORCID iD: 0000-0002-2374-2045
2022 (English)In: Post-transcriptional gene regulation / [ed] Erik Dassi, Humana Press, 2022, 3, , p. 8p. 355-362Chapter in book (Refereed)
Abstract [en]

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.

Place, publisher, year, edition, pages
Humana Press, 2022, 3. , p. 8p. 355-362
Series
Methods in Molecular Biology (MIMB), ISSN 1064-3745, E-ISSN 1940-6029 ; 2404
Keywords [en]
Epitranscriptomics, MeRIP-seq or m6A-seq, METTL3, N6-Methyladenosine
National Category
Cell and Molecular Biology Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-189549DOI: 10.1007/978-1-0716-1851-6_19Scopus ID: 2-s2.0-85118431040ISBN: 9781071618516 (electronic)ISBN: 9781071618509 (print)OAI: oai:DiVA.org:umu-189549DiVA, id: diva2:1611768
Funder
Region VästerbottenKnut and Alice Wallenberg FoundationSwedish Research Council, 2017-01636The Kempe Foundations, 19 0337 Pj, SMK-1766Available from: 2021-11-16 Created: 2021-11-16 Last updated: 2022-03-08Bibliographically approved

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Bhattarai, Devi PrasadAguilo, Francesca

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Bhattarai, Devi PrasadAguilo, Francesca
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Wallenberg Centre for Molecular Medicine at Umeå University (WCMM)Department of Molecular Biology (Faculty of Medicine)
Cell and Molecular BiologyBiochemistry and Molecular Biology

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