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Tet-mediated covalent labelling of 5-methylcytosine for its genome-wide detection and sequencing

Author

Listed:
  • Liang Zhang

    (University of Chicago, 929 E 57th Street, Chicago, Illinois 60637, USA)

  • Keith E. Szulwach

    (Emory University School of Medicine, 615 Michael Street, Atlanta, Georgia 30322, USA)

  • Gary C. Hon

    (Ludwig Institute for Cancer Research, UCSD, Moores Cancer Center, and Institute of Genome Medicine, University of California, San Diego School of Medicine, 9500 Gilman Drive, La Jolla, California 92093-0653, USA)

  • Chun-Xiao Song

    (University of Chicago, 929 E 57th Street, Chicago, Illinois 60637, USA)

  • Beomseok Park

    (University of Illinois at Chicago, 845 West Taylor Street, Chicago, Illinois 60606, USA)

  • Miao Yu

    (University of Chicago, 929 E 57th Street, Chicago, Illinois 60637, USA)

  • Xingyu Lu

    (University of Chicago, 929 E 57th Street, Chicago, Illinois 60637, USA)

  • Qing Dai

    (University of Chicago, 929 E 57th Street, Chicago, Illinois 60637, USA)

  • Xiao Wang

    (University of Chicago, 929 E 57th Street, Chicago, Illinois 60637, USA)

  • Craig R. Street

    (Emory University School of Medicine, 615 Michael Street, Atlanta, Georgia 30322, USA)

  • Huiping Tan

    (Emory University School of Medicine, 615 Michael Street, Atlanta, Georgia 30322, USA)

  • Jung-Hyun Min

    (University of Illinois at Chicago, 845 West Taylor Street, Chicago, Illinois 60606, USA)

  • Bing Ren

    (Ludwig Institute for Cancer Research, UCSD, Moores Cancer Center, and Institute of Genome Medicine, University of California, San Diego School of Medicine, 9500 Gilman Drive, La Jolla, California 92093-0653, USA)

  • Peng Jin

    (Emory University School of Medicine, 615 Michael Street, Atlanta, Georgia 30322, USA)

  • Chuan He

    (University of Chicago, 929 E 57th Street, Chicago, Illinois 60637, USA)

Abstract

5-methylcytosine is an epigenetic mark that affects a broad range of biological functions in mammals. The chemically inert methyl group prevents direct labelling for subsequent affinity purification and detection. Therefore, most current approaches for the analysis of 5-methylcytosine still have limitations of being either density-biased, lacking in robustness and consistency, or incapable of analysing 5-methylcytosine specifically. Here we present an approach, TAmC-Seq, which selectively tags 5-methylcytosine with an azide functionality that can be further labelled with a biotin for affinity purification, detection and genome-wide mapping. Using this covalent labelling approach, we demonstrate high sensitivity and specificity for known methylated loci, as well as increased CpG dinucleotide coverage at lower sequencing depth as compared with antibody-based enrichment, providing an improved efficiency in the 5-methylcytosine enrichment and genome-wide profiling.

Suggested Citation

  • Liang Zhang & Keith E. Szulwach & Gary C. Hon & Chun-Xiao Song & Beomseok Park & Miao Yu & Xingyu Lu & Qing Dai & Xiao Wang & Craig R. Street & Huiping Tan & Jung-Hyun Min & Bing Ren & Peng Jin & Chua, 2013. "Tet-mediated covalent labelling of 5-methylcytosine for its genome-wide detection and sequencing," Nature Communications, Nature, vol. 4(1), pages 1-10, June.
    • Handle: RePEc:nat:natcom:v:4:y:2013:i:1:d:10.1038_ncomms2527
    DOI: 10.1038/ncomms2527
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