Developing\na Ready-to-Use Lipid Nanoparticle Technology\nfor Nucleic Acid Delivery Based on Deep Eutectic Solvents

Microfluidic\ntechnology has emerged as a prevalent tool\nto produce\nlipid nanoparticles (LNPs) for nucleic acid delivery. However, its\nwide-ranging application is hindered by specialized, costly equipment\nand consumables. Herein, a ready-to-use lipid nanoparticle (RULNP)\ntechnology employing deep eutectic solvents (DESs) was developed.\nThe DES, consisting of fructose and glycerol ([Fru][Gly]), was able\nto dissolve lipids and nucleic acids, facilitating the formation of\nRULNPs by simple physical mixing and hydrating. This innovative approach\ncircumvents the high costs and organic solvents associated with microfluidic\nmethods and offers flexibility in preparation techniques, accommodating\nvarious application scenarios. RULNPs exhibited physicochemical properties\nand plasmid DNA (pDNA) or RNA delivery efficacy comparable to those\nof LNPs. Mechanistic studies revealed that RULNPs achieved superior\ncellular uptake compared with LNPs despite exhibiting limited endosomal\nescape capabilities. Collectively, the DES-based RULNP system presents\na rapid and straightforward method for LNP production, potentially\nrevolutionizing nucleic acid delivery.