Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Irvine, CA, USA.
Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Methods Mol Biol. 2021;2282:159-169. doi: 10.1007/978-1-0716-1298-9_10.
The major challenge for RNAi-based therapy is the fabrication of the delivery system that meet the requirement of clinical applicability. Liposome-derived nanoparticles (NPs) are one of the best investigated systems for in vivo siRNA delivery. In the recent years, we have successfully redesigned the conventional cationic liposomes into Liposome/Protamine/hyaluronic acid (LPH) NPs and Lipid-Calcium-Phosphate (LCP) NPs in order to increase the in vivo gene silencing effect and reduce the toxicity. Here we describe the preparation of LPH and LCP NPs loaded with siRNA, and characterization analysis including size distribution, trapping efficiency, and in vivo activity. This protocol could be used for in vivo delivery of siRNA to target genes in cancer cells.
基于 RNAi 的治疗的主要挑战是制造满足临床适用性要求的输送系统。脂质体衍生的纳米颗粒 (NPs) 是体内 siRNA 递送研究最多的系统之一。近年来,我们成功地将传统阳离子脂质体重新设计成脂质体/鱼精蛋白/透明质酸 (LPH) NPs 和脂质-钙-磷酸盐 (LCP) NPs,以提高体内基因沉默效果并降低毒性。在这里,我们描述了负载 siRNA 的 LPH 和 LCP NPs 的制备,并对其进行了包括粒径分布、包封效率和体内活性在内的特征分析。该方案可用于将 siRNA 递送至癌细胞中的靶基因。