Computer-intelligent customized CHA-graphene oxide-DNA circuit for multiplex miRNA detection and accurate cancer cell...

In the analysis of cancer molecular subtypes, miRNAs are important tumor markers. Although artificially designed DNA circuits have been widely used and exhibited high sensitivity in miRNA detection, the drawbacks of low efficiency and lack of high throughput restrict their practical applications. Herein, we propose a fluorescent biosensor assisted with computer-intelligent customization technology, enabling ultrasensitive, multiplex, and simultaneous detection of miRNA-21 and miRNA-30a in solution and cells. Computer-intelligent customization technology is used to precisely output DNA circuit elements, while graphene oxide quenches fluorescence and serves as a carrier for transporting DNA circuits into cells. When the target miRNA is present, the catalytic hairpin assembly reaction is triggered, resulting in fluorescence signal amplification. Under optimal conditions, miRNAs (miRNA-21 and miRNA-30a) can be detected in the range of 20 aM-200 nM within 30 min, with the detection limits of 12.46 and 10.64 aM, respectively. This work provides significant theoretical guidance and practical value for early cancer diagnosis, treatment and subtype identification.