A worm-like nucleic acid nanostructure for gene delivery and endosomal escape via ClC3 ion exchanger

Nanoparticle-based gene delivery can enable therapeutic applications with lower cytotoxicity than viral vectors, but its efficacy is often hampered by endosomal entrapment. We present a nucleic acid nanotechnology approach to circumvent this delivery bottleneck by adsorbing therapeutic nucleic acids (DNA, small interfering RNA, microRNA, or messenger RNA) to a gold-polydopamine nanoworm template, thereby assembling a three-dimensional worm-like nucleic acid nanostructure. Devoid of cationic groups, lipids, or mechanical stimuli, this nanostructure naturally activates the chloride voltage-gated channel 3 (ClC3) ion exchanger in endosomes given its worm-like shape; in turn, ClC3 mediates endosomal H⁺ and Cl^- accumulation and eventual membrane rupture for cytosolic release, contributing to robust endosomal escape with a correlation coefficient <0.2 between the nanostructure and endosomes. We showcase in vitro gene regulation for primary macrophage polarization and mesenchymal stromal cell differentiation, ex vivo programmable mesenchymal stromal cell-based therapy for kidney fibrosis, and in vivo hepatocyte delivery for treating liver injury. Our versatile nucleic acid nanostructure will empower safe and effective gene therapies.