Differential efficacy of DOTAP enantiomers for siRNA delivery in vitro.
Differential efficacy of DOTAP enantiomers for siRNA delivery in vitro.
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DOI：
10.1016/j.ijpharm.2012.04.017
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发表时间：
2012-07-01
影响因子：
5.8
通讯作者：
Lee, Robert J.
中科院分区：
文献类型：
--
作者：
Terp, Megan Cavanaugh;Bauer, Finn;Sugimoto, Yasuro;Yu, Bo;Brueggemeier, Robert W.;Lee, L. James;Lee, Robert J.
DOTAP作为外消旋混合物，是阳离子脂质和广泛使用的转染试剂。在这项研究中，DOTAP的立体化学结构对转染效率的影响进行了评估在体外。在lipoplex制剂中使用外消旋和对映体纯的DOTAP以将siRNA递送至MCF-7细胞，靶向芳香酶。在50 nM siRNA浓度和脂质与RNA电荷比为4和5时，发现DOTAP的R对映体比S-和外消旋剂表现更好。此外，在IOnM siRNA浓度和3的电荷比下，R-lipoplex制剂使芳香酶沉默约50%，而S和外消旋制剂没有引起显著的靶下调。使用膜模拟对脂质堆积的差异进行建模。结果显示，当与胆固醇组合时，纯R-DOTAP和S-DOTAP对映体相对于外消旋DOTAP分别具有105%和115%的脂质密度。这些发现表明脂质手性在基于脂质的siRNA递送系统的未来发展中的重要作用。
DOTAP, as a racemic mixture, is a cationic lipid and a widely used transfection reagent. In this study, the effect of DOTAP’s stereochemical structure on transfection efficiency was evaluated in vitro. Racemic and enantiomerically pure DOTAP were used in lipoplex formulations to deliver siRNA to MCF-7 cells, targeting the aromatase enzyme. At the 50nM siRNA concentration and lipid-to-RNA charge ratios of 4 and 5, the R enantiomer of DOTAP was found to perform better than either the S- and the racemic agent. In addition, at 10 nM siRNA concentration and a charge ratio of 3, the R- lipoplex formulation silenced aromatase by ~ 50% whereas the S and racemic formulations caused no significant target downregulation. Differences in lipid packing were modeled using membrane simulations. The results showed that, when combined with cholesterol, pure R-DOTAP and S-DOTAP enantiomers had 105% and 115% of lipid density relative to racemic DOTAP, respectively. These findings suggest an important role of lipid chirality in future development of lipid based siRNA delivery systems.
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DOI：
10.1083/jcb.141.4.905
发表时间：
1998-05-18
期刊：
The Journal of cell biology
影响因子：
--
作者：
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通讯作者：
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影响因子：
5.8
作者：
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DOI：
10.1016/s0927-7757(01)00543-x
发表时间：
2001-07-15
影响因子：
5.2
作者：
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通讯作者：
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作者：
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影响因子：
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