Issue 11, 2017

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

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

Supplementary files

Article information

Article type
Edge Article
Submitted
23 Aug 2017
Accepted
04 Sep 2017
First published
04 Sep 2017
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2017,8, 7443-7447

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, 8, 7443 DOI: 10.1039/C7SC03685J

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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