CRISPR-driven biosensing is developing rapidly, but its signal reporters are still hard to balance cost and performance, which has greatly limited its development. Herein, we use HCR reaction to generate DNA-CuNCs as the reporter for CRISPR/Cas12a-based assay, successfully developing a rapid detection platform for disease markers. The hairpins for HCR reaction rich A and T bases at the rings and sticky ends that can produce poly (AT-TA) as the template of DNA-CuNCs. On the other hand, the activator of the CRISPR/Cas12a system contained aptamers that recognize the target. As the activator preferentially binds to the target, CRISPR/Cas12a remains silent, and the HCR reaction generates poly (AT-TA) dsDNA to fabricate fluorescent DNA-CuNCs. Inversely, the Cas12a is triggered by activator in the absence of the target to trans-cleaved the components of HCR, resulting in the failure of DNA-CuNCs formation. By using HCR and modified fluorescence-stable DNA-CuNCs, we constructed a highly sensitive platform that can detect disease markers in one-pot reaction within 1 hour, providing a new detection protocol for point-of-care tests.

CRISPR/Cas12a-based biosensor enabled by HCR-dependent DNA-copperclusters for disease markers detection

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