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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

作者： Wang, Zemiao 1 ; Zhang, Feng 1 ; Fang, Ling 1 ; Chen, Fengping 1 ; Yang, Weijuan 1 ; Wang, Zongwen 1 ;

作者机构： 1.Fujian Agr & Forestry Univ, Coll Plant Protect, State Key Lab Ecol Pest Control Fujian & Taiwan Cr, Fuzhou 350002, Fujian, Peoples R China

2.Fujian Agr & Forestry Univ, Key Lab Biopesticide & Chem Biol, Minist Educ, Fuzhou 350002, Fujian, Peoples R China

3.Fujian Acad Agr Sci, Inst Qual Stand & Testing Technol Agroprod, Fujian Key Lab Agroprod Qual & Safety, Fuzhou 350003, Peoples R China

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

页码：

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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