Ultrasensitive Photoelectrochemical Detection of MicroRNA on Paper by Combining a Cascade Nanozyme-Engineered Biocatalytic Precipitation Reaction and Target-Triggerable DNA Motor

Developing efficient strategies for sensitive detection of microRNAs, the noncoding bioactive molecules and well-established biomarkers, has aroused great interests due to its great potential values in genetic and pathological analyses. Herein, a highly selective and disposable paper-based photoelectrochemical (PEC) sensor was rationally designed for sensing microRNA based on simple self-assembly of a target-triggerable DNA motor and nanozyme-catalyzed multistage biocatalytic precipitation reaction. Specifically, a brand-new type II heterojunction of TiO2-CeO2 nanotubes decorated with carbon fiber paper (CFP) was first prepared, which gave an enhanced photoreactive surface and realized fast electron transport and extraction, markedly accelerating photoelectric conversion efficiency of the sensor. For achieving target detection, cascade nanozyme centers of the CeO2 and Au nanoparticles modified by cyclodextrin were drafted, greatly decreasing the photocurrent intensity and achieving an ultralow background signal. With target introduction, the DNA motor was activated and automatically moved along the predesigned route driven by an endonuclease cleavage reaction, resulting in more substrate probe digestion and nanozyme release from CFP. Consequently, the repressive inner enhancement mechanism was gradually renewed with constant advancement of the enzymatic reaction and walker probe walking progressively, eventually allowing multiple enzymatic factor output in each target import. As a proof-of-concept application, the developed PEC sensor successfully performed detection of miRNA-141, showing a low detection limit of 0.6 fM, and was further applied to real sample bioassays with satisfying results. This work proposes promising strategies to boost the catalytic cascade DNA-motor adhibition in biological samples analysis and also exhibits potential capability in detection of other targets.