Design of lipid nanoparticles for in vitro and in vivo delivery of plasmid DNA
Design of lipid nanoparticles for in vitro and in vivo delivery of plasmid DNA
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DOI：
10.1016/j.nano.2016.12.014
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发表时间：
2017-05-01
影响因子：
5.4
通讯作者：
Cullis, Pieter R.
中科院分区：
文献类型：
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作者：
Kulkarni, Jayesh A.;Myhre, Johnathan Layne;Cullis, Pieter R.
脂质纳米颗粒（LNPs）含有二硬脂酰磷脂酰胆碱（dsc）和可电离的氨基脂质，如二烯油基甲基-4二甲氨基丁酸酯（DLin-MC3-DMA）是体内有效的siRNA递送载体。在这里，我们探讨了类似LNP系统作为质粒DNA （pDNA）转染试剂的效用。结果表明，用不饱和pc代替dsc和用相关脂质DLin-KC2-DMA代替DLin-MC3-DMA可以在体外产生高效的HeLa细胞转染试剂。此外，这些配方在多种哺乳动物细胞系中表现出优异的转染特性，在原代细胞培养中转染效率接近90%。这些转染水平等于或大于脂质体所达到的水平，毒性大大降低。最后，将LNPeGFP显微注射到鸡胚胎的肢体芽中，报告基因得到了强烈的表达。综上所述，含有可电离氨基脂质的LNP系统在体内和体外都是一种高效、无毒的基因表达pDNA传递系统。(C) 2016 Elsevier Inc.版权所有。
Lipid nanoparticles (LNPs) containing distearoylphosphatidlycholine (DSPC), and ionizable amino-lipids such as dilinoleylmethyl-4dimethylaminobutyrate (DLin-MC3-DMA) are potent siRNA delivery vehicles in vivo. Here we explore the utility of similar LNP systems as transfection reagents for plasmid DNA (pDNA). It is shown that replacement of DSPC by unsaturated PCs and DLin-MC3-DMA by the related lipid DLin-KC2-DMA resulted in highly potent transfection reagents for HeLa cells in vitro. Further, these formulations exhibited excellent transfection properties in a variety of mammalian cell lines and transfection efficiencies approaching 90% in primary cell cultures. These transfection levels were equal or greater than achieved by Lipofectamine, with much reduced toxicity. Finally, microinjection of LNPeGFP into the limb bud of a chick embryo resulted in robust reporter-gene expression. It is concluded that LNP systems containing ionizable amino lipids can be highly effective, non-toxic pDNA delivery systems for gene expression both in vitro and in vivo. (C) 2016 Elsevier Inc. All rights reserved.