Effect of Mg ions on efficiency of gene electrotransfer and on cell electropermeabilization
Effect of Mg ions on efficiency of gene electrotransfer and on cell electropermeabilization
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DOI：
10.1016/j.bioelechem.2010.04.001
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发表时间：
2010-10-01
影响因子：
5
通讯作者：
Pavlin, Mojca
中科院分区：
文献类型：
--
作者：
Haberl, Sasa;Miklavcic, Damijan;Pavlin, Mojca
基因电转移是一种很有前途的非病毒方法，它可以通过电脉冲将DNA转移到活细胞中。然而，有许多参数决定基因电转移效率。基因电转移的步骤之一是DNA与细胞膜的相互作用。电穿孔介质中的二价阳离子可以影响DNA锚定到细胞膜上，并由此影响基因电转移效率。本文报道了不同浓度的Mg ~（2+）对小分子碘化丙啶的电渗透、基因电转移和细胞活力的影响。我们还使用TOTO-1染料来可视化不同[Mg]的DNA-细胞膜相互作用。为此目的，我们使用不同的电穿孔介质与增加[Mg]。我们的研究表明，更高的[Mg]导致碘化丙啶的更高的电透化和更高的活力，同时导致较低的基因电转移效率。因为我们在使用较高[Mg]时在细胞表面观察到较高的TOTO-1标记的DNA，所以我们认为Mg 2+离子可以以在施加电脉冲期间不能进入细胞的强度在细胞表面结合DNA，这可能导致较低的基因转染。也可能有其他机制参与，因为有许多步骤的基因电转移上的Mg 2+离子可以有一个effecton. Our结果还意味着，膜通透性的变化是不够的，一个有效的基因电转移。(C)2010 Elsevier B. V.保留所有权利。
Gene electrotransfer is a promising nonviral method that enables DNA to be transferred into living cells with electric pulses. However, there are many parameters that determine gene electrotransfer efficiency. One of the steps involved in gene electrotransfer is interaction of DNA with the cell membrane. Divalent cations in the electroporative media can influence the anchoring of DNA to the cell membrane and by that gene electrotransfer efficiency. Here we report the effect of different concentrations of Mg2+ on electropermeabilization for small molecule (propidium iodide), gene electrotransfer and viability of the cells. We also used TOTO-1 dye to visualize DNA-cell membrane interaction for different [Mg]. For this purpose, we used different electroporative media with increasing [Mg]. Our study shows that higher [Mg] lead to higher electropermeabilization for propidium iodide and higher viability, while causing lower gene electrotransfer efficiency. Because we observed higher TOTO-1 labeled DNA at cell surface when using higher [Mg] we suggest that Mg2+ ions can bind DNA at cell surface at such strength that cannot pass into the cell during application of electric pulses, which can lead to lower gene transfection. There may also be other mechanisms involved, since there are many steps of gene electrotransfer on which Mg2+ ions can have an effect on. Our results also imply that membrane permeability changes are not sufficient for an efficient gene electrotransfer. (C) 2010 Elsevier B.V. All rights reserved.