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

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 wi...

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Published in:Chemical science (Cambridge) 2017, Vol.8 (11), p.7443-7447
Main Authors: Liu, Chaoxing, Wang, Yafen, Yang, Wei, Wu, Fan, Zeng, Weiwu, Chen, Zonggui, Huang, Jinguo, Zou, Guangrong, Zhang, Xiong, Wang, Shaoru, Weng, Xiaocheng, Wu, Zhiguo, Zhou, Yu, Zhou, Xiang
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Language:English
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Summary: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.
ISSN:2041-6520
2041-6539
DOI:10.1039/c7sc03685j