Insights into the mechanisms of electromediated gene delivery and application to the loading of giant vesicles with negatively charged macromolecules
Insights into the mechanisms of electromediated gene delivery and application to the loading of giant vesicles with negatively charged macromolecules
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深入了解电介导基因传递的机制及其在带负电大分子负载巨囊泡中的应用
作者：
Thomas Portet;Cyril Favard;Justin Teissi'e;David S. Dean;M. Rols
我们提出了质粒DNA电转移到磷脂酰胆碱巨型单层囊泡（GUVs）中的实验结果。我们的观察表明，直接进入是电转移的主要机制。我们还对guv内的DNA浓度增量进行了定量分析，发现我们的实验数据可以很好地用一个简单的理论模型来描述，其中DNA的进入主要是由电泳驱动的。我们的理论框架允许预测转移DNA的数量作为电场参数的函数，从而为一种新的方法铺平了道路，这种方法不仅可以高效地封装DNA，还可以将任何带负电荷的大分子封装到guv中。
We present experimental results regarding the electrotransfer of plasmid DNA into phosphatidylcholine giant unilamellar vesicles (GUVs). Our observations indicate that a direct entry is the predominant mechanism of electrotransfer. A quantitative analysis of the DNA concentration increments inside the GUVs is also performed, and we find that our experimental data are very well described by a simple theoretical model in which DNA entry is mostly driven by electrophoresis. Our theoretical framework allows for the prediction of the amount of transferred DNA as a function of the electric field parameters, and thus paves the way towards a novel method for encapsulating with high efficiency not only DNA, but any negatively charged macromolecules into GUVs.
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