Kanamori Takashi, Ohzeki Hiroki, Masaki Yoshiaki, Ohkubo Akihiro, Takahashi Mari, Tsuda Kengo, Ito Takuhiro, Shirouzu Mikako, Kuwasako Kanako, Muto Yutaka, Sekine Mitsuo, Seio Kohji
Education Academy of Computational Life Sciences, Tokyo Institute of Technology, 4259 Nagatsuta, Midoriku, Yokohama 226-8501 (Japan).
Chembiochem. 2015 Jan 2;16(1):167-76. doi: 10.1002/cbic.201402346. Epub 2014 Dec 2.
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 dU(MBF) 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).
我们开发了一种荧光开启型探针，其包含荧光核苷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))，用于通过利用DNA三链体形成来检测单链DNA或RNA。荧光测量结果表明，含有dU(MBF)的探针比含有dU(BF)的探针实现了更好的荧光增强。核磁共振和荧光分析表明，三链体形成时荧光强度增加，部分原因是3-甲基苯并呋喃与尿嘧啶环之间的键发生了构象变化。此外，有人认为3-甲基苯并呋喃环周围的微环境有助于荧光增强。此外，我们还开发了一种结合滚环扩增与含有dU(MBF)的寡核苷酸探针的三链体诱导荧光增强来检测RNA的方法。
