CRISPR/Cas12a-Responsive Hydrogel Biosensing Platform Based on an Aggregation-induced Emission Metal–Organic Framework Nanozyme for Sensitive and Reliable Detection of Foodborne Pathogens

Programmable CRISPR/Cas12a has emerged as a promising and powerful tool for the construction of biosensors, yet the challenges persist regarding the signal transduction efficiency, sensitivity, and multisignal outputs. In this work, a metal-organic framework (MOF) nanozyme with an aggregation-induced emission (AIE) property and peroxidase (POD)-like activity was synthesized by encapsulating hemin into a Zr-based MOF using an in situ encapsulation strategy, named AIE MOFzyme. The prepared AIE MOFzyme showed an enhanced fluorescent property via the restriction of the intramolecular motion effect for boosting the significant fluorescent response and a highly efficient POD-like nanozyme activity for activating visual colorimetric response. Benefiting from these merits, AIE MOFzyme, as a difunctional signal probe, was embedded into a DNA hydrogel for the construction of a CRISPR-responsive AIE MOFzyme hydrogel platform (abbreviated as Cas12a-H-AIE). In this platform, the activated CRISPR/Cas12a system led to DNA hydrogel phase transitions to release AIE MOFzymes, triggering highly efficient fluorescent and colorimetric signal amplification. Importantly, the CRISPR-responsive AIE MOFzyme hydrogel platform for the detection of <i>Salmonella enterica</i> (<i>S. enterica</i>) showed a high sensitivity, which was superior to that of numerous existing methods. This work provided new insight for the design of multifunctional and programmable nanozyme hydrogels, which may also offer guidance for the development of novel CRISPR/Cas12a-based biosensors.