Review: engineering CRISPR nanoparticle hybrid platforms for biofilm-associated antimicrobial resistance: delivery barriers, resistance reprogramming, and translational challenges

Abstract

Abstract Antimicrobial resistance represents a critical global health threat that outpaces the development of new antibiotics. Biofilms impose a multifaceted barrier, comprising an extracellular polymeric substance matrix of polysaccharides, proteins, and extracellular DNA, that limits antibiotic penetration and facilitates horizontal gene transfer of resistance determinants. This review adopts a biofilm-centered engineering framework to evaluate clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 nanoparticle hybrids as an emerging precision antimicrobial strategy, analyzing the sequential barriers of matrix penetration, bacterial envelope traversal, intracellular cargo release, and resistance-network reprogramming that determine therapeutic success. Lipid-nanoparticle-mediated CRISPR-Cas9 delivery and gold-nanoparticle CRISPR hybrids have been explored for antimicrobial co-delivery applications; precise, matched quantitative comparisons between nanoparticle platforms specific to biofilm CRISPR delivery remain limited in the primary literature. By precisely targeting quorum-sensing regulators ( lasR, rhlR, luxS ), resistance genes ( mcr-1, mecA, blaNDM-1, blaCTX-M-15 ), and biofilm structural genes ( icaA, icaD, bap, csgD ), this platform is designed to convert biofilm from an obstacle into a targetable interface. However, clinical translation faces major hurdles, including manufacturing complexity, immunogenicity, off-target risks, regulatory ambiguity between nanomedicine and gene therapy frameworks, and long-term biosafety concerns such as horizontal gene transfer. This review concludes that CRISPR-nanoparticle hybrids offer a transformative platform for precision antimicrobial intervention; however, no clinical trials currently exist for this specific application, and coordinated advances in nanoparticle engineering, safety characterization, and regulatory science are essential for clinical deployment against recalcitrant, multidrug-resistant biofilm infections.