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

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Fluorogenic labeling and single-base resolution analysis of 5-formylcytosine in DNA
Chemical Science ( IF 7.6 ) Pub Date : 2017-09-04 00:00:00 , DOI: 10.1039/c7sc03685j
Chaoxing Liu _{1,

2,

3,

4,

5} , Yafen Wang _{1,

2,

3,

4,

5} , Wei Yang _{1,

2,

3,

4,

5} , Fan Wu _{1,

2,

3,

4,

5} , Weiwu Zeng _{1,

2,

3,

4,

5} , Zonggui Chen _{5,

6,

7,

8} , Jinguo Huang _{1,

2,

3,

4,

5} , Guangrong Zou _{1,

2,

3,

4,

5} , Xiong Zhang _{1,

2,

3,

4,

5} , Shaoru Wang _{1,

2,

3,

4,

5} , Xiaocheng Weng _{1,

2,

3,

4,

5} , Zhiguo Wu _{5,

6,

7,

8} , Yu Zhou _{5,

6,

7,

8} , Xiang Zhou _{1,

2,

3,

4,

5}

Affiliation

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.

中文翻译：

DNA中5-甲酰基胞嘧啶的荧光标记和单碱基拆分分析

在表观遗传功能中起重要作用的5-甲酰基胞嘧啶（5fC）已在许多相关领域引起广泛关注。在这里，我们演示了通过选择性地使试剂与5fC反应生成分子内环化核碱基，对5fC进行荧光检测和单碱基拆分分析的新设计。生成的产物带有类似的苯并噻唑-亚氨基香豆素骨架，具有很高的荧光强度，使我们能够定性和定量地检测γ射线照射的小牛胸腺DNA中的5fC部分。此外，在5fC中失去环外4-氨基会导致dATP在聚合酶延伸过程中通过与生成的核碱基的碱基配对而掺入，这有助于我们分析单链和双链寡核苷酸中的5fC位点。我们的Sanger和Illumina测序结果显示了5fC单碱基分辨率分析的巨大潜力。希望类似的设计可以用于更多的检测目标。

更新日期：2017-09-20

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