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Identification of a Selective DNA Ligase for Accurate Recognizing and Ultrasensitive Quantifying N6-Methyladenosine in RNAs at One-Nucleotide Resolution

Abstract

N6-methyladenosine (m6A) is the most frequently post-transcriptional modification in RNAs, which plays a critical role in biological processes. The m6A functions remain largely unexplored due to lack of highly sensitive methods to quantitatively determine the m6A modification fraction at the precise location. Here, we firstly reveal that T3 DNA ligase has significant selectivity against m6A modification. On the basis of the new finding, we establish an ultrasensitive quantitation assay for accurate determining m6A at one-nucleotide resolution in RNAs. With the proposed assay, as low as 4 fM RNA-containing m6A can be determined and the selectivity is up to 54.1-fold to discriminate m6A against unmodified adenosine (A). The sensitivity has been improved about 106-fold, so that the proposed method can be successfully employed to accurately determine the m6A in real biological samples, even in low abundant RNAs.

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Publication details

The article was received on 10 Dec 2017, accepted on 15 Feb 2018 and first published on 15 Feb 2018


Article type: Edge Article
DOI: 10.1039/C7SC05233B
Citation: Chem. Sci., 2018, Accepted Manuscript
  • Open access: Creative Commons BY license
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    Identification of a Selective DNA Ligase for Accurate Recognizing and Ultrasensitive Quantifying N6-Methyladenosine in RNAs at One-Nucleotide Resolution

    W. liu, J. Yan, Z. Zhang, H. Pian, C. Liu and Z. Li, Chem. Sci., 2018, Accepted Manuscript , DOI: 10.1039/C7SC05233B

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