Yuan Jing, Deng Zhiwei, Liu Hui, Li Xiufang, Li Jianbing, He Yao, Qing Zhihe, Yang Yanjing, Zhong Shian
College of Chemistry and Chemical Engineering, Central South University, Changsha, 410083, PR China.
School of Chemistry and Food Engineering, Changsha University of Science and Technology, Changsha, 410114, PR China.
ACS Sens. 2019 Jun 28;4(6):1648-1653. doi: 10.1021/acssensors.9b00482. Epub 2019 Jun 19.
The precise detection of extracellular ATP, although a challenging task, is of great significance for understanding the related cell processes. Herein, we developed a ratiometric DNA nanoswitch by employing a DNA tweezer and split aptamer. The nanoswitch is composed of three specially designed ssDNA strands, namely, the central strands O1, O2, and O3. This nanoswitch can be anchored on the cell membrane by cholesterol labeled at the 3' end of O3. Initially, the DNA tweezer adopts an open state, separating the dual fluorophores and giving rise to a low FRET (fluorescence resonance energy transfer) ratio. The presence of ATP induces the binding of the two split aptamers to alter the structure of the nanoswitch from the open state to the closed state, bringing the donor and the acceptor closer together and generating high FRET efficiency. The results demonstrated that the ratiometric DNA nanoswitch can be applied for quantitative analysis and real-time monitoring of the changes in extracellular ATP. We believe that the cell surface-anchored DNA nanoswitch has promising prospects for use as a powerful tool for the understanding of different ATP-related physiological activities.
细胞外ATP的精确检测虽然是一项具有挑战性的任务，但对于理解相关细胞过程具有重要意义。在此，我们通过使用DNA镊子和分裂适体开发了一种比率型DNA纳米开关。该纳米开关由三条特别设计的单链DNA组成，即中心链O1、O2和O3。这种纳米开关可以通过标记在O3 3'端的胆固醇锚定在细胞膜上。最初，DNA镊子处于开放状态，分离双荧光团并产生低荧光共振能量转移（FRET）比率。ATP的存在诱导两个分裂适体结合，使纳米开关的结构从开放状态变为闭合状态，使供体和受体靠得更近并产生高FRET效率。结果表明，比率型DNA纳米开关可用于细胞外ATP变化的定量分析和实时监测。我们相信，细胞表面锚定的DNA纳米开关作为理解不同ATP相关生理活动的强大工具具有广阔的应用前景。
