Fusogenic RNA Nanomodules for Fusion‐Mediated and Multiplexed siRNA Delivery

Abstract

Lipid nanoparticles (LNPs) are state‐of‐the‐art siRNA carriers but are limited by modest RNA loading, inefficient endosomal escape, and short‐lived silencing due to burst release at escape. To overcome these constraints, we developed lipid‐layered core RNA‐assembled nanomodules (L‐CRAMs) that couple a self‐assembled RNA core with fusogenic lipids to enable sustained, high‐capacity siRNA delivery. L‐CRAMs release siRNA gradually, exhibit fusion‐mediated intracellular delivery with limited endo‐lysosomal sequestration, and produce robust silencing across single and multiplexed targets in vitro. Compared with MC3‐LNPs, L‐CRAMs prolonged intracellular gene silencing. Following systemic administration in mice, L‐CRAMs induced potent, durable suppression of the clinically relevant liver gene APOC3, accompanied by reductions in serum triglycerides (TG) and triglyceride‐rich lipoproteins (TRL). By integrating ultra‐high RNA payloads, fusogenic uptake, and programmable multi‐gene targeting, L‐CRAMs provide a versatile, long‐acting RNA interference platform that addresses key limitations of existing LNP systems and advances therapeutic RNA delivery.