Universal Design of Structure-Switching Aptamers with Signal Reporting Functionality
| Autor: | Hualong Gao, Shuo Wang, Yang Huang, Yu Han, Jiaoxing Zhao, Xinhui Lou, Xiao Cheng, Yi Xiao |
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| Rok vydání: | 2019 |
| Předmět: |
Aptamer
Deoxyribozyme Sequence (biology) Biosensing Techniques Computational biology 010402 general chemistry G-quadruplex 01 natural sciences Signal Analytical Chemistry Domain (software engineering) Cocaine Limit of Detection Benzothiazoles Fluorescent Dyes Peroxidase Structure (mathematical logic) Estradiol Chemistry Sulfadimethoxine 010401 analytical chemistry Thrombin DNA Catalytic Mercury Aptamers Nucleotide 0104 chemical sciences G-Quadruplexes Spectrometry Fluorescence Biocatalysis Nucleic Acid Conformation Colorimetry Biosensor |
| Zdroj: | Analytical Chemistry. 91:14514-14521 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.9b03368 |
| Popis: | Structure-switching aptamers (SSAs) offer a promising recognition element for sensor development. However, the generation of SSAs via in vitro aptamer selection technologies or postselection engineering is challenging. Inspired by the two-domain structure of antibodies, we have devised a simple, universal strategy for engineering aptamers into SSAs with signal reporting functionality. These constructs consist of a "constant" domain, comprising a split DNAzyme G-quadruplex (G4) region for signal transduction, and a "variable" domain, comprising an aptamer sequence capable of specific target binding. In the absence of target, the G4-SSA construct folds into a parallel G4 structure with high peroxidase catalytic activity. Target binding disrupts the G4 structure, resulting in low enzymatic activity. We demonstrate that this change in DNAzyme activity enables sensitive and specific colorimetric detection of diverse targets including Hg2+, thrombin, sulfadimethoxine, cocaine, and 17β-estradiol. G4-SSAs can also achieve label-free fluorescence detection of various targets using a specific G4-binding dye. We demonstrate that diverse aptamers can be readily engineered into G4-SSA constructs independent of target class, binding affinity, aptamer length, or structure. This design strategy could broadly extend the power, accessibility, and utility of numerous SSA-based biosensors. |
| Databáze: | OpenAIRE |
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