Department of Chemistry, Graduate School of Science, Tohoku University , Sendai 980-8578, Japan.
J Am Chem Soc. 2016 Aug 3;138(30):9397-400. doi: 10.1021/jacs.6b05554. Epub 2016 Jul 26.
We have developed a new fluorescent sensing probe for double-stranded RNA (dsRNA) by integrating thiazole orange (TO) as a base surrogate into triplex-forming PNA. Our probe forms the thermally stable triplex with the target dsRNA at acidic pH; and the triplex formation is accompanied by the remarkable light-up response of the TO unit. The binding of our probe to the target dsRNA proceeds very rapidly, allowing real-time monitoring of the triplex formation. Importantly, we found the TO base surrogate in our probe functions as a universal base for the base pair opposite the TO unit in the triplex formation. Furthermore, the TO unit is significantly more responsive for the fully matched dsRNA sequence compared to the mismatch-containing sequences, which enables the analysis of the target dsRNA sequence at the single-base pair resolution. The binding and sensing functions of our probe are described for the development of fluorescent probes applicable to sensing biologically relevant dsRNA.
我们通过将噻唑橙（TO）整合到三链体形成的 PNA 中作为碱基替代物，开发了一种用于双链 RNA（dsRNA）的新型荧光传感探针。我们的探针在酸性 pH 下与靶 dsRNA 形成热稳定的三链体；并且三链体的形成伴随着 TO 单元的显著点亮响应。探针与靶 dsRNA 的结合非常迅速，允许实时监测三链体的形成。重要的是，我们发现我们的探针中的 TO 碱基替代物在三链体形成中充当与 TO 单元相对的碱基的通用碱基。此外，与包含错配的序列相比，TO 单元对完全匹配的 dsRNA 序列的响应更显著，这使得能够以单碱基对分辨率分析靶 dsRNA 序列。我们的探针的结合和传感功能可用于开发适用于感测生物相关 dsRNA 的荧光探针。
