Dynamic 3D DNA Rolling Walkers via Directional Movement on a Lipid Bilayer Supported by Au@Fe3O4 Nanoparticles for Sensitive Detection of MiRNA-16

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Abstract
Currently reported polyfluorene-based fluorescence detection usually shows high background signal and low detection sensitivity. This work developed a novel three-dimensional (3D) DNA rolling walker via directional movement on a lipid bilayer (LB) supported by Au@Fe3O4 nanoparticles (NPs) in a polyfluorene-based fluorescence system so that it could achieve significantly improved detection sensitivity and almost zero-background signal detection for miRNA-16. First, the carboxyl-functionalized poly[(9,9-dioctylfluorenyl-2,7-diyl)-co-(1,4-benzo-{2,1',3}-thiadazole)] polymer nanoparticles (c-PFBT PNPs) covalently bonded with amino-labeled single-strand CP and further hybridized with single-strand AP to prepare AP-CP-coupled c-PFBT PNP probes. Meanwhile, Au@Fe3O4 NPs were developed as efficient fluorescence quenchers and served as the matrix for assembling the LB. The resulting Au@Fe3O4@LB assembled cholesterol-labeled orbital DNA L1 and L2 and further assembled hairpins H1 and AP-CP-coupled c-PFBT PNP probes to construct DNA nanomachines. Then, the target miRNA-16 was introduced to initiate the rolling circle amplification (RCA) reaction and form dynamic DNA rolling walkers, thus releasing single-strand CP-coupled c-PFBT PNP probes. The magnetic separation effect of Au@Fe3O4 NPs made it possible to detect the fluorescence signal from the released probes, thus achieving almost zero-background signal detection for miRNA-16 with a low detection limit of 95 aM. The flexible interfaces provided by the LB endowed the DNA rolling walkers with high binding efficiency and low derailment probability, thus achieving significantly improved detection sensitivity. The developed strategy provided an attractive polyfluorene-based fluorescence platform with high-sensitivity and low-background signals.
Funding Information
  • Natural Science Foundation of Chongqing Municipality (cstc2018jcyjAX0693)
  • National Natural Science Foundation of China (21775122, 21775124)

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