Controlling the Fluorescence of Benzofuran-Modified Uracil Residues in Oligonucleotides by Triple-Helix Formation
Controlling the Fluorescence of Benzofuran-Modified Uracil Residues in Oligonucleotides by Triple-Helix Formation
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DOI：
10.1002/cbic.201402346
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发表时间：
2015-01-02
期刊：
影响因子：
3.2
通讯作者：
Seio, Kohji
中科院分区：
文献类型：
--
作者：
Kanamori, Takashi;Ohzeki, Hiroki;Seio, Kohji
我们开发了含有荧光核苷5-(苯并呋喃-2-基)脱氧尿苷(Du(BF))或5-(3-甲基苯并呋喃-2-基)脱氧尿苷(Du(MBF))的荧光开启探针，用于利用DNA三链形成检测单链DNA或RNA。荧光测量表明，含dUMBF的探针比含Du(BF)的探针具有更好的荧光增强效果。核磁共振和荧光分析表明，在三链形成时荧光强度增加，部分原因是3-甲基苯并呋喃和尿嘧啶环之间的键的构象变化。此外，3-甲基苯并呋喃环周围的微环境对荧光增强也有贡献。此外，我们发展了一种滚动循环扩增结合含Du(MBF)的三链诱导荧光增强的寡核苷酸探针检测RNA的方法。
We developed fluorescent turn-on probes containing a fluorescent nucleoside, 5-(benzofuran-2-yl) deoxyuridine (dU(BF)) or 5-(3-methylbenzofuran-2-yl) deoxyuridine (dU(MBF)), for the detection of single-stranded DNA or RNA by utilizing DNA triplex formation. Fluorescence measurements revealed that the probe containing dUMBF achieved superior fluorescence enhancement than that containing dU(BF). NMR and fluorescence analyses indicated that the fluorescence intensity increased upon triplex formation partly as a consequence of a conformational change at the bond between the 3-methylbenzofuran and uracil rings. In addition, it is suggested that the microenvironment around the 3-methylbenzofuran ring contributed to the fluorescence enhancement. Further, we developed a method for detecting RNA by rolling circular amplification in combination with triplex-induced fluorescence enhancement of the oligonucleotide probe containing dU(MBF).