Buffering properties of cationic polymethacrylates are not the only key to successful gene delivery
Buffering properties of cationic polymethacrylates are not the only key to successful gene delivery
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DOI：
10.1021/bm034438d
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发表时间：
2004-03-01
影响因子：
6.2
通讯作者：
Schacht, E
中科院分区：
文献类型：
--
作者：
Dubruel, P;Christiaens, B;Schacht, E
最近，我们发现含咪唑侧基团(HYMIMMA)或酸官能团(MA)的聚甲基丙烯酸酯(HYMIMMA)或酸官能团(MA)具有类似于聚乙烯亚胺的缓冲性能，但不能转染Cos-1细胞，而只含叔胺的聚合物(DMAEMA)确实能转染Cos-1细胞(Dubrel，P.et al.欧元。J·帕姆。SCI。2003、18(3-4)、211-220)。在目前的工作中，我们调查了细胞内化和/或亚细胞定位的差异在多大程度上与转染活性的差异有关。因此，我们合成了一系列含伯胺功能的聚甲基丙烯酸酯类化合物，用于俄勒冈绿荧光探针的偶联。用含有荧光素衍生物(5-氨甲基)荧光素盐酸盐的胺标记含有酸性功能的聚合物。研究表明，MA和基于HYMIMMA的复合体的内体释放可能是Cos-1细胞基因转移过程中的限制步骤。
Recently, we have shown that polymethacrylates containing imidazole side groups (HYMIMMA) or acid functions (MA), which have similar buffering properties as polyethyleneimine, were not able to transfect Cos-1 cells, whereas polymers containing only tertiary amines (DMAEMA) do transfect Cos-1 cells (Dubruel, P. et al. Eur. J. Pharm. Sci. 2003, 18 (3-4), 211-220). In the present work, we investigated to what extent the differences in transfection activity are related to differences in cellular internalization and/or subcellular localization. Therefore, we synthesized a series of polymethacrylates containing primary amine functions, used for the coupling of the fluorescent Oregon Green probe. The polymers containing acid functions were labeled with an amine containing fluorescein derivative (5-aminomethyl)fluorescein hydrochloride. It is demonstrated that the endosomal release of the MA and HYMIMMA-based complexes might be the limiting step in the gene transfer process in Cos-1 cells.