Fluorogenic labeling and single-base resolution analysis of 5-formylcytosine in DNA. Issue 11 (19th September 2017)

• Record Type: Journal Article
• 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
• Is Part Of: Chemical science. Volume 8:Issue 11(2017)
• Page Start: 7443
• Page End: 7447
• Publication Date: 2017-09-19
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/c7sc03685j
• Languages: English
• ISSNs: 2041-6520

Abstract

Abstract : Simultaneous fluorogenic switch-on detection and single-base resolution analysis of 5fC through yielding an intramolecular cyclization nucleobase has been presented. 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.