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Epitranscriptomics pp 69–79Cite as

High-Resolution Mapping of N 6-Methyladenosine Using m6A Crosslinking Immunoprecipitation Sequencing (m6A-CLIP-Seq)

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1870))

Abstract

N 6-Methyladenosine, an abundant chemical modification in mRNA, plays crucial roles in regulating gene expression and biological processes. Research on m6A and its functions has progressed rapidly in the past few years, aided substantially by advances in high-throughput sequencing-based methods to profile m6A along the transcriptome. We present here a protocol for m6A crosslinking immunoprecipitation sequencing (m6A-CLIP-seq), which profiles m6A on mRNA at high resolution from as little as 1 μg of poly(A)-selected mRNA.

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Acknowledgements

We thank Caroline Vissers, Boxuan Zhao, and Allen Zhu for helpful comments. P.J.H. is supported by the NIH Medical Scientist National Research Service Award T32 GM007281. C.H. is supported by National Institutes of Health grants GM071440 and HG008935. C.H. is an investigator of the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. Department of Chemistry and Institute for Biophysical Dynamics; Howard Hughes Medical Institute, The University of Chicago, Chicago, IL, USA

    Phillip J. Hsu & Chuan He

  2. Medical Scientist Training Program/Committee on Immunology, The University of Chicago, Chicago, IL, USA

    Phillip J. Hsu

  3. Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL, USA

    Chuan He

Authors
  1. Phillip J. Hsu
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  2. Chuan He
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Corresponding author

Correspondence to Chuan He .

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Editors and Affiliations

  1. Department of Pathology, Yale University School of Medicine, New Haven, CT, USA

    Narendra Wajapeyee

  2. Yale Cancer Center, Yale University School of Medicine, New Haven, CT, USA

    Romi Gupta

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Hsu, P.J., He, C. (2019). High-Resolution Mapping of N 6-Methyladenosine Using m6A Crosslinking Immunoprecipitation Sequencing (m6A-CLIP-Seq). In: Wajapeyee, N., Gupta, R. (eds) Epitranscriptomics. Methods in Molecular Biology, vol 1870. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8808-2_5

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  • DOI: https://doi.org/10.1007/978-1-4939-8808-2_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8807-5

  • Online ISBN: 978-1-4939-8808-2

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