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Fluorogenic labeling and single-base resolution analysis of 5-formylcytosine in DNA

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Abstract

5-Formylcytosine (5fC), which plays an important role in epigenetic functions, has received widespread attention in many related fields. Here, we demonstrate a new design for both the fluorogenic switch-on detection and single-base resolution analysis of 5fC through selectively reacting a reagent with 5fC to yield an intramolecular cyclization nucleobase. The generated product, bearing a similar benzothiazole-iminocoumarin scaffold, is highly fluorescent and enables us to qualitatively and quantitatively detect 5fC moieties in γ-irradiated calf thymus DNA. Additionally, losing the exocyclic 4-amino group in 5fC causes the incorporation of dATP through base pairing with the generated nucleobase during polymerase extension, which helped us to analyze the 5fC sites in both single- and double-stranded oligonucleotides. Our Sanger and Illumina sequencing results show great potential in single-base resolution analysis of 5fC. It is hopeful that a similar design may be used for more detection targets.

Graphical abstract: Fluorogenic labeling and single-base resolution analysis of 5-formylcytosine in DNA

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Publication details

The article was received on 23 Aug 2017, accepted on 04 Sep 2017 and first published on 04 Sep 2017


Article type: Edge Article
DOI: 10.1039/C7SC03685J
Citation: Chem. Sci., 2017, Advance Article
  • Open access: Creative Commons BY license
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    Fluorogenic labeling and single-base resolution analysis of 5-formylcytosine in DNA

    C. Liu, Y. Wang, W. Yang, F. Wu, W. Zeng, Z. Chen, J. Huang, G. Zou, X. Zhang, S. Wang, X. Weng, Z. Wu, Y. Zhou and X. Zhou, Chem. Sci., 2017, Advance Article , DOI: 10.1039/C7SC03685J

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