Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, Shaanxi Normal University, Xi'an 710062, China.
Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry & Chemical Engineering, Shaanxi Normal University, Xi'an 710062, China.
Anal Chem. 2021 Jan 26;93(3):1333-1341. doi: 10.1021/acs.analchem.0c02637. Epub 2020 Dec 21.
As a recently identified higher-order quadruplex (G4) structure, the G4 dimer possesses unique structure and biological functions. In this work, we found accidentally that two tandem PW17 (one known G4-forming DNA) sequences can fold into a stable G4 dimer, and the G4 dimer can enhance dramatically the fluorescence intensity of thioflavin T (ThT). The G4 dimer/ThT fluorescence intensity is about ninefold that of the corresponding G4 monomer/ThT. Meanwhile, compared with the common G4/ThT system, G4 dimer/ThT exhibited more stable fluorescence emission in the media with various concentrations of Na and K. On the basis of these findings, G4 dimer/ThT was used as a fluorescence indicator to construct one arched DNA probe for label-free detection of DNA. By incorporating a G4 dimer sequence in amplified products, we further designed one rolling circle amplification-based biosensing strategy to show the utility of this G4 dimer/ThT fluorescence indicator. This study demonstrates that dimeric G4 is an effective nucleic acid scaffold for lighting up ThT, showing promising applications in a label-free bioassay.
作为最近发现的一种高阶四联体(G4)结构,G4 二聚体具有独特的结构和生物学功能。在这项工作中,我们偶然发现两个串联的 PW17(一个已知的 G4 形成 DNA)序列可以折叠成稳定的 G4 二聚体,并且 G4 二聚体可以显著增强硫黄素 T(ThT)的荧光强度。G4 二聚体/ThT 的荧光强度大约是相应 G4 单体/ThT 的九倍。同时,与常见的 G4/ThT 系统相比,G4 二聚体/ThT 在具有各种浓度的 Na 和 K 的介质中表现出更稳定的荧光发射。基于这些发现,G4 二聚体/ThT 被用作荧光指示剂来构建一种无标记的 DNA 检测弧形 DNA 探针。通过在扩增产物中并入 G4 二聚体序列,我们进一步设计了一种基于滚环扩增的生物传感策略,以展示这种 G4 二聚体/ThT 荧光指示剂的实用性。这项研究表明,二聚体 G4 是一种有效的核酸支架,可点亮 ThT,在无标记生物测定中具有广阔的应用前景。