Fluorogenic labeling and single-base resolution analysis of 5-formylcytosine in DNA.
Fluorogenic labeling and single-base resolution analysis of 5-formylcytosine in DNA.
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DNA 中 5-甲酰胞嘧啶的荧光标记和单碱基分辨率分析
DOI：
10.1039/c7sc03685j
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发表时间：
2017-11-01
期刊：
影响因子：
8.4
通讯作者：
Zhou X
中科院分区：
文献类型：
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作者：
Liu C;Wang Y;Yang W;Wu F;Zeng W;Chen Z;Huang J;Zou G;Zhang X;Wang S;Weng X;Wu Z;Zhou Y;Zhou X
同时荧光开关检测和单碱基分辨率分析5 fC通过产生一个分子内环化核碱基已提交。5-甲酰基胞嘧啶（5 fC）在表观遗传功能中发挥着重要作用，在许多相关领域受到广泛关注。在这里，我们展示了一种新的设计，通过选择性地与5 fC反应的试剂，以产生分子内环化核碱基的荧光开关检测和单碱基分辨率分析5 fC。所产生的产物具有类似的苯并噻唑-亚氨基香豆素骨架，具有高荧光性，使我们能够定性和定量检测γ辐照小牛胸腺DNA中的5 fC部分。此外，在5 fC中失去环外4-氨基导致dATP通过在聚合酶延伸期间与生成的核碱基碱基配对而掺入，这有助于我们分析单链和双链寡核苷酸中的5 fC位点。我们的桑格和Illumina测序结果显示了5 fC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。
Simultaneous fluorogenic switch-on detection and single-base resolution analysis of 5fC through yielding an intramolecular cyclization nucleobase has been presented. 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.
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影响因子：
14.9
作者：
Madugundu GS;Cadet J;Wagner JR
通讯作者：
Wagner JR
RNA 中 5-甲基胞嘧啶氧化产物的形成和测定
影响因子：
8.4
作者：
Huang W;Lan MD;Qi CB;Zheng SJ;Wei SZ;Yuan BF;Feng YQ
通讯作者：
Feng YQ
影响因子：
15
作者：
Hardisty RE;Kawasaki F;Sahakyan AB;Balasubramanian S
通讯作者：
Balasubramanian S
影响因子：
16.8
作者：
Raiber, Eun-Ang;Murat, Pierre;Chirgadze, Dimitri Y.;Beraldi, Dario;Luisi, Ben F.;Balasubramanian, Shankar
通讯作者：
Balasubramanian, Shankar
影响因子：
21.8
作者：
通讯作者：
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