CRISPR/Cas12a-Sheared ZIF-Based Heterojunction to Allow Polarity-Switchable Photoelectrochemical and Nanozyme-Enabled Colorimetric Dual-Modal Biosensing

Abstract

Modulating the migration of interfacial carriers in heterojunctions is critical for driving the signal response of high-performance optical biosensors. In this study, a polarity-switchable photoelectrochemical (PEC) and nanozyme-enabled colorimetric dual-modal biosensor is designed to modulate the interfacial carrier migration of the zeolitic imidazolate framework (ZIF)-based heterojunction by exploiting stem-loop DNA and the CRISPR/Cas12a system. Specifically, ZIF-hemin (ZIF-Hemin) is assembled at the CdSe/NH₂-rGO interface via stem-loop DNA to form a ZIF-based heterojunction. Stem-loop DNA with a reinforcing rib effect enhances binding and accelerates the interfacial carrier migration of the heterojunction. In the presence of the target Cry1Ab, the CRISPR/Cas12a system is activated to shear the ZIF-based heterojunction, resulting in the disintegration of the heterojunction and the disappearance of interfacial carrier migration. At this point, ZIF-Hemin is released from the CdSe/NH₂-rGO interface, with the photocurrent switching from the anode to the cathode. Meanwhile, due to its rich accessible active sites, the released ZIF-Hemin nanosheet shows high peroxidase-like catalytic activity and generates colorimetric signals. The dual-modal biosensor demonstrates excellent performance in selectivity and sensitivity, with low detection limits of 0.05 pg mL^–1 (PEC) and 0.4 pg mL^–1 (colorimetric). This work provides a general strategy to improve the performance of optical biosensors by modulating the migration of interfacial carriers in heterojunctions.