A surface-mediated hydrogel patch for localized CRISPR genome editing in inflammatory wound therapy.

Abstract

The NLRP3 inflammasome is a central driver of inflammatory tissue injury and delayed repair in bacteria-infected wounds, yet local genome editing in such lesions remains limited by nonspecific release and off-target exposure. Here, we report a pH-gated surface-mediated hydrogel patch for infection-responsive delivery of CRISPR-Cas9 ribonucleoprotein (RNP) complexes. Rather than embedding the cargo within the bulk matrix, polymer-coated NLRP3-targeting RNPs are reversibly anchored on the hydrogel surface, enabling localized presentation under physiological conditions and rapid detachment in the acidic microenvironment of Staphylococcus aureus (S. aureus)-infected wounds. In vitro studies demonstrated an NLRP3 editing efficiency of 57% in J774A.1 cells and 69% in L929 cells surpassing or matching the performance of the commercial transfection reagent Lipofectamine CRISPRMAX (50% in J774A.1; comparable in L929). In a murine full-thickness skin defect model with S. aureus infection, topical administration of the patch achieved >20% NLRP3 editing efficiency in wound tissue, accompanied by a statistically significant reduction in inflammatory cell infiltration. Consequently, this pH-responsive surface anchoring and release strategy establishes a novel paradigm for localized, precision gene therapy and holds strong translational promise for the treatment of chronic inflammatory wounds.