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小分子干扰RNA(siRNA)和质粒DNA(pDNA)与阳离子脂质体的复合:脂质体复合物制备中的重要方面。

Complexation of siRNA and pDNA with cationic liposomes: the important aspects in lipoplex preparation.

作者信息

Barichello José Mario, Ishida Tatsuhiro, Kiwada Hiroshi

机构信息

Department of Pharmacokinetics and Biopharmaceutics, Subdivision of Biopharmaceutical Sciences, Institute of Health Biosciences, The University of Tokushima, Tokushima, Japan.

出版信息

Methods Mol Biol. 2010;605:461-72. doi: 10.1007/978-1-60327-360-2_32.

Abstract

In the last two decades, cationic liposomes have been investigated as vehicles for nucleic acids [plasmid DNA (pDNA) and small interfering RNA (siRNA)] delivery in vitro and in vivo. The formation of cationic liposomes-nucleic acids complexes, termed lipoplexes, depends on a number of experimental variables. The quality of the nucleic acid and the cationic liposome as well as the selection of diluents for diluting the concentrated stocks strongly affect the resulting lipoplexes and their efficiency of gene-expression or gene-silencing effect following transfection. In addition, the molar ratio of cationic lipid nitrogen (N) to siRNA or pDNA phosphate (P) (N/P ratio) influences the final characteristics of the lipoplexes, such as size, surface zeta potential, and reproducibility, thereby reflecting their efficiency following transfection. The methods presented in this chapter could be helpful to obtain reliable and reproducible lipoplexes and experimental results.

摘要

在过去二十年中,阳离子脂质体已被研究作为核酸(质粒DNA(pDNA)和小干扰RNA(siRNA))在体外和体内递送的载体。阳离子脂质体 - 核酸复合物(称为脂质体复合物)的形成取决于许多实验变量。核酸和阳离子脂质体的质量以及用于稀释浓缩储备液的稀释剂的选择强烈影响所得的脂质体复合物及其转染后基因表达或基因沉默效果的效率。此外,阳离子脂质氮(N)与siRNA或pDNA磷酸(P)的摩尔比(N/P比)影响脂质体复合物的最终特性,如大小、表面zeta电位和重现性,从而反映其转染后的效率。本章介绍的方法可能有助于获得可靠且可重复的脂质体复合物和实验结果。

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