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Efficient Mrna Delivery Using Lipid Nanoparticles Modified with Fusogenic Coiled-Coil Peptides.

Nanoscale(2023)

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摘要
Gene delivery has great potential in modulating protein expression in specific cells to treat diseases. Such therapeutic gene delivery demands sufficient cellular internalization and endosomal escape. Of various nonviral nucleic acid delivery systems, lipid nanoparticles (LNPs) are the most advanced, but still, are very inefficient as the majority are unable to escape from endosomes/lysosomes. Here, we develop a highly efficient gene delivery system using fusogenic coiled-coil peptides. We modified LNPs, carrying EGFP-mRNA, and cells with complementary coiled-coil lipopeptides. Coiled-coil formation between these lipopeptides induced fast nucleic acid uptake and enhanced GFP expression. The cellular uptake of coiled-coil modified LNPs is likely driven by membrane fusion thereby omitting typical endocytosis pathways. This direct cytosolic delivery circumvents the problems commonly observed with the limited endosomal escape of mRNA. Therefore fusogenic coiled-coil peptide modification of existing LNP formulations to enhance nucleic acid delivery efficiency could be beneficial for several gene therapy applications. Regular nanoparticles delivery efficiency is hindered by endosome entrapment. By modifying lipid-nanoparticles with lipopeptide CPE4 and pretreating the cells with complimentary CPK4, coiled-coil peptides induced significantly enhanced mRNA delivery in various cell lines mediated by membrane fusion.
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