Overcoming the nuclear barrier:: Cell cycle independent nonviral gene transfer with linear polyethylenimine or electroporation
Overcoming the nuclear barrier:: Cell cycle independent nonviral gene transfer with linear polyethylenimine or electroporation
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DOI：
10.1006/mthe.2001.0509
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发表时间：
2002-01-01
影响因子：
12.4
通讯作者：
Wagner, E
中科院分区：
文献类型：
--
作者：
Brunner, S;Fürtbauer, E;Wagner, E
在许多情况下，非病毒颗粒介导的基因传递高度依赖于转染细胞的细胞周期状态。在这里，我们比较了线性聚乙烯亚胺（PEI）的颗粒介导传递和分支PEI的电穿孔DNA物理转移以及脂肪转染在细胞周期不同阶段的转染能力。与其他颗粒介导的递送方法（使用Lipofectamine或支链PEI）相比，线性PEI在G1和S/G2转染的HeLa细胞之间的基因转移中仅导致很小的差异（在1个对数单位内）。平行转染（脂肪转染或分支PEI）导致G1和S/G2转染的细胞之间荧光素酶表达差异2至3个对数单位。电穿孔基因转移也几乎没有细胞周期依赖性，表现出完全不同的表达动力学。电穿孔后3小时报告基因的表达已经非常高，在所有细胞周期阶段报告基因的表达水平大致相同。我们认为，与其他测试的DNA传递系统相比，DNA电穿孔和DNA转染与线性PEI颗粒具有更好的核输入特性。
In many cases, nonviral particle-mediated gene delivery is highly dependent on the cell cycle status of transfected cells. Here we compare particle-mediated delivery with linear polyethylenimine (PEI) and physical transfer of DNA by electroporation with branched PEI and lipofection for their ability to transfect cells at different stages of the cell cycle. In contrast to other particle-mediated delivery methods (using Lipofectamine or branched PEI) linear PEI led to only small differences (within 1 log unit) in gene transfer between HeLa cells transfected in G1 and those in S/G2. Parallel transfections (lipofection or branched PEI) resulted in 2 to > 3 log-unit differences in luciferase expression between cells transfected in G1 and S/G2. Gene transfer by electroporation also revealed hardly any cell cycle dependence and displayed completely different expression kinetics. Reporter gene expression is already very high 3 hours after electroporation with roughly the same level of reporter gene expression in all cell cycle phases. We suggest that DNA electroporation and DNA transfection with linear PEI particles have improved nuclear import characteristics relative to the other tested DNA delivery systems.