Development of folding-based and signal-amplified fluorescence aptasensors using G-quadruplex quinclorac aptamer variants engineered via exonuclease digestion and circular dichroism spectroscopy

文献类型： 外文期刊

第一作者： Wang, Zemiao

作者： Wang, Zemiao;Zhang, Feng;Fang, Ling;Chen, Fengping;Yang, Weijuan;Wang, Zongwen;Wang, Zemiao;Zhang, Feng;Fang, Ling;Chen, Fengping;Yang, Weijuan;Wang, Zongwen;Fang, Ling

作者机构：

关键词： Phosphorodiamidate morpholino oligomer; Recycling amplification; Exonuclease III; Exonuclease I; N -methyl mesoporphyrin IX

期刊名称：SENSORS AND ACTUATORS B-CHEMICAL （ 影响因子：8.0； 五年影响因子：7.0 ）

ISSN：

年卷期： 2024 年 415 卷

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收录情况： SCI

摘要： The lack of inherent structure -switching functionality and inadequate affinity in aptamers hinders the construction and application of aptasensors. While exonuclease digestion -based strategies have been employed to engineer small -molecule aptamers with the structure -switching functionality and construct dual-exonuclease (exonuclease I and exonuclease III) digestion -based aptasensors, such studies targeting G-quadruplex (G4) aptamers remain scarce. This study focused on the exonuclease digestion and circular dichroism spectroscopy analysis of a G4 quinclorac (QNC) aptamer, Qapt-51, intending to design a functionalized QNC aptamer and a universal signal amplification strategy for constructing folding -based and sensitive dual-exonuclease digestionbased aptasensors, respectively. The exonuclease digestion results demonstrated that QNC-binding Qapt-51 inhibited the dual-exonuclease digestion. Combining the results from the circular dichroism spectroscopy, we further proposed a digestion mechanism for Qapt-51 involving a G4 conformational switch. Inspired by this mechanism, a functionalized Qapt-51 variant (Qapt-45A) was rationally designed to construct a simple and rapid folding -based fluorescence aptasensor. Additionally, we developed an exonuclease III- and phosphorodiamidate morpholino oligomer-assisted fluorescence signal amplification strategy, which can work in the dual-exonuclease digestion system. Leveraging this strategy integrated with the dual-exonuclease digestion of another Qapt-51 variant (AQ), a homogeneous signal -amplified fluorescence aptasensor was constructed to sensitively detect QNC without requiring the functionalized aptamers. These two aptasensors achieved detection limits of 560 ng/ mL and 8.5 ng/mL for QNC. They successfully detected QNC in rice and river water samples with recoveries of 108.6%-87.5%. We hope that the successful development of these two aptasensors can provide valuable insights and references for the broader construction of G4 aptamer-based sensing systems.

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