LAUSR

In this study, a novel polyhistidine (pHis)-incorporated lipid nanoparticle (pHis/LNP) was developed for the delivery of therapeutic globotriaosylceramide (Gb3) synthase siRNAs using a microfluidic device with pHis as a biocompatible method of endosome escape. To inhibit the expression of Gb3 synthase, six siRNAs against Gb3 synthase were designed and an optimal siRNA sequence was selected. Selected Gb3 synthase siRNA was incorporated into pHis/LNP to prepare a spherical siRNA pHis/LNP with a size of 62.5 ± 1.9 nm and surface charge of -13.3 ± 4.2 mV. The pHis/LNP successfully protected siRNAs from degradation in 50% serum condition for 72 h. Prepared pHis/LNP exhibited superior stability for 20 days and excellent biocompatibility for A549 cells. After treatment with fluorescence-labeled LNPs, dotted fluorescent signals were co-localized with Lysotracker in cells with LNPs, whereas strong and diffused fluorescence intensity was observed in cells with pHis/LNPs probably due to successful endosomal escape. The extent of Gb3 synthase gene silencing by siRNA pHis/LNP was greatly improved (6.0-fold) compared to that by siRNA/LNP. Taken together, considering that the fabricated siRNA pHis/LNP exhibited excellent biocompatibility and superior gene silencing activity over conventional LNP, these particles can be utilized for the delivery of a wide range of therapeutic siRNAs. This article is protected by copyright. All rights reserved.