Terminal-overhang-restricted CRISPR logic nanodevice for precision cancer therapy.

The limited precision of controlled drug release remains a major challenge for precision cancer therapy. Herein, a protein and nucleic acid dual-biomarker-responsive CRISPR AND logic nanodevice, termed LogiCas, is developed by integrating APE1 and miRNA-21 as dual inputs for precise tumor therapy. Leveraging the stringent conformational matching required for Cas12a activation, LogiCas utilizes the terminal redundancy restriction of the split activator to establish a dual-lock, stepwise activation mechanism consisting of APE1-mediated pre-unlocking and miRNA-21-mediated terminal activation, enforcing tightly gated control over Cas12a trans-cleavage. The activated Cas12a further cleaves Ce6-BHQ3 to trigger Ce6-mediated photodynamic therapy (PDT), generating reactive oxygen species (ROS) that in turn drive the nuclear-to-cytoplasmic translocation of APE1, contributing to an APE1-mediated positive feedback effect. In a nude mouse model, LogiCas achieves tumor-site-specific drug release, demonstrating robust antitumor efficacy with ~91% tumor inhibition, without obvious systemic toxicity. This work provides a new design strategy for integrating protein and nucleic acid biomarkers across molecular levels to enable CRISPR-mediated precision therapeutic activation.