Polyethylenimine-mediated gene delivery: a mechanistic study
Polyethylenimine-mediated gene delivery: a mechanistic study
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DOI：
10.1002/jgm.173
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发表时间：
2001-03-01
影响因子：
3.5
通讯作者：
Danos, O
中科院分区：
文献类型：
--
作者：
Kichler, A;Leborgne, C;Danos, O
乙烯亚胺聚合物（PEI）属于用于将质粒DNA递送到哺乳动物细胞中的最有效的阳离子化合物家族之一。即使在不存在内体溶解剂如融合肽或氯喹的情况下也获得高转染效率，这与大多数其他阳离子聚合物相反。它已被假设，PEI的效率是由于它的能力，以缓冲endosome.Methods调查的重要性，在PEI介导的DNA transfer过程中，我们使用质子泵抑制剂，如巴弗洛霉素Al和concanamycin A的酸化的内体。此外，我们测试是否PEI是能够不稳定的天然膜本身在中性或酸性pH值通过执行红细胞裂解assays.Results PEI介导的转染存在下，巴弗洛霉素铝导致在7-74倍的报告基因的表达减少，这取决于所使用的细胞系。相比之下，单阳离子脂质DOTAP的效率在药物存在下没有显著改变。此外，目前的数据表明，PEI不能不稳定的红细胞膜，即使在酸性pH值，和PEI，复合或不与DNA，可以增加转染效率的阳离子聚合物，聚赖氨酸，当在同一时间加入到cells.Conclusions的PEI的转染效率部分依赖于他们的能力，捕获质子被转移到内体在其酸化。此外，PEI能够将显著量的DNA递送到细胞中，并且参与转基因表达的DNA复合物在4小时内从内体逃逸。版权所有（C）2001约翰威利父子有限公司
Background Ethylenimine polymers (PEIs) belong to one of the most efficient family of cationic compounds for delivery of plasmid DNA into mammalian cells. The high transfection efficiencies are obtained even in the absence of endosomolytic agents such as fusogenic peptides or chloroquine, which is in contrast to most of the other cationic polymers. It has been hypothesized that the efficiency of PEI is due to its capacity to buffer the endosomes.Methods To investigate the importance of the acidification of endosomes during PEI-mediated DNA transfer we used proton pump inhibitors such as bafilomycin Al and concanamycin A. Moreover, we tested whether PEI is able to destabilize natural membranes per se at neutral or acidic pH by performing erythrocyte lysis assays.Results PEI-mediated transfection in the presence of bafilomycin Al resulted in a 7-74-fold decrease in reporter gene expression depending on the cell line used. In contrast, the efficiency of the monocationic lipid, DOTAP, was not importantly altered in the presence of the drug. Furthermore, the present data show that PEI cannot destabilize erythrocyte membranes, even at acidic pH, and that PEI, complexed or not to DNA, can increase the transfection efficiency of the cationic polymer, polylysine, when added at the same time to the cells.Conclusions The transfection efficiency of PEIs partially relies on their ability to capture the protons which are transferred into the endosomes during their acidification. In addition, PEI is able to deliver significant amounts of DNA into cells and the DNA complexes involved in the expression of the transgene escape within 4 h from the endosomes. Copyright (C) 2001 John Wiley & Sons, Ltd.