Quadrilateral Nucleic Acid Frame-Accelerating DNAzyme Walker Kinetics for Biosensing Based on Host-Guest Recognition-Enhanced Electrochemiluminescence.

Depending on the reaction between walkers and tracks, DNA walker is able to output signals continuously, which has attracted great attention from the bioanalytical community. Therefore, how to improve its reaction kinetics for efficient signal readout is of great significance. Herein, a quadrilateral DNAzyme walker was fabricated by colocalizing one walker and three DNA tracks in the quadrilateral nucleic acid frame to form a reaction unit (abbreviated as qDNA walker). Impressively, in contrast to the common free DNAzyme walker, the reaction kinetics of the qDNA walker was 2.3 times faster, which could achieve microRNA detection within 30 min. Meanwhile, an electrochemiluminescence (ECL) emitter of anthracene-cucurbituril supramolecular nanocrystals (Ant-CB SNCs) was obtained based on the self-assembly of cucurbituril (CB, host molecule) and anthracene (Ant, guest molecule). Benefiting from the host-guest recognition effect, the prepared Ant-CB SNCs exhibited enhanced ECL efficiency due to the supramolecular interaction between CB and Ant, which could inhibit vibration and rotation of the Ant molecules. We defined this new enhanced ECL phenomenon as "host-guest recognition-enhanced ECL." As a proof of concept, an ECL biosensor for microRNA-21 (miRNA-21) was constructed by combining the high-efficiency DNAzyme walker and the advanced ECL emitter of Ant-CB SNCs, which showed a linear range from 50 aM to 50 pM with a low limit of detection (11 aM), highlighting the great potential in clinical diagnosis.