Abstract
Antibiotic-resistant bacteria cause more than one million deaths annually worldwide. The rapid evolution and gene exchange among pathogens often render new antibiotic drugs ineffective soon after deployment, underscoring the urgent need for alternative therapeutic strategies. Nanoscale silver is well known for its innate bacteriostatic and bactericidal activity but typically requires high concentrations for efficacy that causes toxicities and limits broader clinical applications. To address this, we introduce programmable self-assembling DNA scaffolds that template, stabilize, and spatially organize multiple copies of monodisperse silver nanoclusters (DNA-AgNCs). These assemblies enhance antimicrobial potency of formulations while also exhibiting intrinsic fluorescence, providing dual functionality for therapeutic and bioimaging applications. Detailed characterization identified DNA-AgNC scaffolds with improved stability and enhanced activity against clinically relevant antibiotic-resistant planktonic ESKAPE pathogens. We also revealed the DNA-AgNC constructs that significantly lowered intracellular infections of human bone cells with Staphylococcus aureus . Collectively, the results highlight spatially organized DNA-AgNCs as a promising modular platform for next-generation antibacterial therapy and real-time bioimaging.