Kinetics-accelerated one-step detection of MicroRNA through spatially localized reactions based on DNA tile self-assembly

The localization of isothermal amplification systems has elicited extensive attention due to the enhanced reaction kinetics when detecting ultra-trace small-molecule nucleic acids. Therefore, the seek for an appropriate localization cargo of spatially confined reactions is urgent. Herein, we have developed a novel approach to localize the catalytic hairpin assembly (CHA) system into the DNA tile self-assembly nanostructure. Thanks to the precise programming and robust probe loading capacity, this strategy achieved a 2.3 x 105-fold higher local reaction concentration than a classical CHA system with enhanced reaction kinetics in theory. From the experimental results, this strategy could reach the reaction plateau faster and get access to a magnified effect of 1.57-6.99 times higher in the linear range of microRNA (miRNA) than the simple CHA system. Meanwhile, this strategy satisfied the demand for the one-step detection of miRNA in cell lysates at room temperature with good sensitivity and specificity. These features indicated its excellent potential for ultra-trace molecule detection in clinical diagnosis and provided new insights into the field of bioassays based on DNA tile self-assembly nanotechnology.

Automated summary

This paper presents a novel approach to localize the catalytic hairpin assembly (CHA) system into the DNA tile self-assembly nanostructure, resulting in higher reaction concentration and enhanced reaction kinetics. Experimental results show that this method achieves faster reaction plateau and amplifies the effect of microRNA (miRNA) 1.57-6.99 times higher compared to the simple CHA system. Moreover, this strategy fulfills the requirement for one-step detection of miRNA in cell lysates at room temperature.