Electro-mediated gene transfer and expression are controlled by the life-time of DNA/membrane complex formation
Electro-mediated gene transfer and expression are controlled by the life-time of DNA/membrane complex formation
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DOI：
10.1002/jgm.1414
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发表时间：
2010-01-01
影响因子：
3.5
通讯作者：
Rols, Marie-Pierre
中科院分区：
文献类型：
--
作者：
Faurie, Cecile;Rebersek, Matej;Rols, Marie-Pierre
背景电穿孔是一种用于将分子转移到细胞和组织中的物理方法。抗肿瘤药物递送的临床应用已经发展。基因电转移的临床试验正在调查中。然而，关于DNA如何进入细胞的知识并不完整。与直接进入细胞质的小分子相比，DNA与质膜形成长寿命的复合物，并以相当长的延迟被转移到细胞质中。我们研究了DNA/结果我们观察到，电脉冲极性和重复频率对细胞膜相互作用和基因表达均有影响，增加脉冲的重复频率。在本研究中获得的结果揭示了存在两类DNA/膜相互作用：（i）一个亚稳态的DNA/膜复合物，DNA可以离开和返回到外部介质和（ii）一个稳定的DNA/膜复合物，其中DNA不能被删除，即使通过施加极性相反的电脉冲。只有属于第二类的DNA导致有效的基因expression.Conclusions的DNA/膜复合物形成的寿命是1秒的顺序，并已考虑到改善协议的电介导的基因传递。版权所有（C）2009约翰威利父子有限公司
Background Electroporation is a physical method used to transfer molecules into cells and tissues. Clinical applications have been developed for antitumor drug delivery. Clinical trials of gene electrotransfer are under investigation. However, knowledge about how DNA enters cells is not complete. By contrast to small molecules that have direct access to the cytoplasm, DNA forms a long lived complex with the plasma membrane and is transferred into the cytoplasm with a considerable delay.Methods To increase our understanding of the key step of DNA/membrane complex formation, we investigated the dependence of DNA/membrane interaction and gene expression on electric pulse polarity and repetition frequency.Results We observed that both are affected by reversing the polarity and by increasing the repetition frequency of pulses. The results obtained in the present study reveal the existence of two classes of DNA/membrane interaction: (i) a metastable DNA/membrane complex from which DNA can leave and return to external medium and (ii) a stable DNA/membrane complex, where DNA cannot be removed, even by applying electric pulses of reversed polarity. Only DNA belonging to the second class leads to effective gene expression.Conclusions The life-time of DNA/membrane complex formation is of the order of 1 s and has to be taken into account to improve protocols of electro-mediated gene delivery. Copyright (C) 2009 John Wiley & Sons, Ltd.