DNA nanomachine-based regenerated sensing platform: a novel electrochemiluminescence resonance energy transfer strategy for ultra-high sensitive detection of microRNA from cancer cells.

The construction of DNA nanomachines holds great significance in the development of DNA nanostructures; however, the real application of nanomachines is still in its early stage. Moreover, one-step regenerated sensing platforms for the detection of biomarkers in the current research remain a practical challenge. Herein, a novel electrochemiluminescence resonance energy transfer (ERET) strategy between Alexa Flour 488 (AF 488), which is a type of small molecule dye, as the donor and CdSe@ZnS quantum dots (QDs) as the acceptor, which easily enter the cells, has been reported and was applied for the construction of a DNA nanomachine-based regenerated biosensor for the ultra-high sensitive determination of cancer cells without any enzyme. First, a dual amplification strategy, including target recycling and signal transformation, was employed to achieve the conversion of a small number of miRNAs into a large amount of universal DNA reporters. Initially, the DNA tweezer was kept in the "off" state with two arms labeled with QDs and AF488, respectively. Second, in the presence of DNA reporters, the tweezer transformed to the "on" state through the hybridization of the reporter DNA and exposed the arms of the tweezer. Simultaneously, QDs and AF488 on the two arms were close enough to generate ERET, which remarkably increased the ECL intensity of the QDs. Impressively, the sensor could be regenerated by a one-step strand displacement and could be cycled for more than seven times. Owing to the dual amplification strategy and the high efficiency of the ERET between the QDs and AF488, the proposed biosensor performs in the linear range from 10 pM to 0.1 fM with a detection limit of 0.03 fM for miRNA determination, and the monitoring of different cancer cells was also achieved. Moreover, the elaborated biosensor can also realize the sensitive detection of Pb2+, which indicates that it can be potentially used for field environmental analysis and monitoring, thus offering a new modular platform for the construction of functional DNA nanomachines in the ultra-high sensitive analysis of promising biomarkers and toxic metals.