Proximity-Dependent Switchable ATP Aptasensors Utilizing a High-Performance FRET Reporter
Proximity-Dependent Switchable ATP Aptasensors Utilizing a High-Performance FRET Reporter
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利用高性能 FRET 报告基因的邻近依赖性可切换 ATP 适体传感器
DOI：
10.1021/acsami.0c15543
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Tao Li
中科院分区：
文献类型：
--
作者：
Qiwei Wang;Tai Fang;Jiao Zheng;Lin Shi;Lili Shi;Tao Li
在这里，荧光分子转子被用来开发一种新型高性能 FRET 系统，该系统具有大斯托克斯位移、高光稳定性和 pH 不敏感性，在邻近依赖性 DNA 适体传感器中显示出巨大的应用前景。合成了两个基于羧基苯并噻唑的分子转子，一旦标记到 DNA 上，就会显示出明亮的绿色和红色荧光。在接近状态下，两种染料之间会发生有效的 FRET，与最常用的 Cy3/Cy5 对相当。如果采用基于萘并噻唑的类似物，也会观察到类似的现象。然后，我们开发的 FRET 对连接到由双分子 i-motif 控制的二聚 DNA 纳米支架中分裂 ATP 适体的两部分。通过这种方式，构建了pH切换的邻近诱导荧光ATP适体传感器，具有良好的灵敏度、选择性和可重构性。此外，通过改变开关单元的接头长度，系统地研究了邻近效应，为DNA邻近反应及其在某些生理过程中的作用提供了新的见解。
Here, fluorescent molecular rotors are employed to develop a new type of high-performance FRET system with large Stokes shift, high photostability, and pH insensitivity, showing great promise for use in proximity-dependent DNA aptasensors. Two carboxylated benzothiazole-based molecular rotors are synthesized, displaying bright green and red fluorescence once labeled to DNA. In the proximity state, an efficient FRET occurs between the two dyes, comparable to that of the most commonly used Cy3/Cy5 pair. Similar phenomena are also observed if naphthothiazole-based analogues are adopted. Our developed FRET pair is then attached to the two parts of a split ATP aptamer in a dimeric DNA nanoscaffold controlled by a bimolecular i-motif. In this way, a pH-switched proximity-induced fluorescent ATP aptasensor is constructed, with good sensitivity, selectivity, and reconfiguration. Furthermore, by altering the linker length of the switching unit, the proximity effect is investigated systematically, providing new insight into DNA proximity reactions and their roles in some physiological processes.