2-(dimethylamino)ethyl methacrylate based (co)polymers as gene transfer agents
2-(dimethylamino)ethyl methacrylate based (co)polymers as gene transfer agents
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DOI：
10.1016/s0168-3659(97)00248-4
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发表时间：
1998-04-30
影响因子：
10.8
通讯作者：
Hennink, WE
中科院分区：
文献类型：
--
作者：
van de Wetering, P;Cherng, JY;Hennink, WE
聚甲基丙烯酸2-（二甲氨基）乙酯（PDMAEMA）是一种水溶性阳离子聚合物，能够通过静电相互作用与DNA结合。在聚合物/质粒比率高于2（w/w）时，形成大小约为0.2 μ m的带正电荷的复合物。使用编码β-半乳糖苷酶的pCMV-lacZ质粒作为报告基因，在细胞培养物（COS-7和OVCAR-3细胞）中评价聚合物/质粒复合物的转染效率。在PDMAEMA/质粒比例为3-5（w/w）时发现最佳转染效率。在这些条件下，3-6%的细胞实际上被转染。与其他阳离子聚合物一样，PDMAEMA具有轻微的细胞毒性。这种活性被聚合物与DNA的复合部分掩盖。观察到聚合物的分子量对转染效率的显著影响。增加的分子量导致转染细胞的数量增加。动态光散射实验表明，高分子量聚合物（M-w>300 kDa）能够有效地凝聚DNA（粒径0.15-0.20 μ m）。相反，当质粒与低分子量PDMAEMA一起孵育时，形成大的复合物（大小为0.5-1.0 μ m）。DMAEMA与甲基丙烯酸甲酯（MMA）、乙氧基三甘醇甲基丙烯酸酯（triEGMA）或N-乙烯基吡咯烷酮（NVP）的共聚物也充当转染剂。与相同分子量的均聚物相比，具有20摩尔%的MMA的共聚物显示出降低的转染效率和显著增加的细胞毒性。与DMAEMA均聚物相比，具有triEGMA（48摩尔%）的共聚物显示出降低的转染效率和降低的细胞毒性，而具有NVP（54摩尔%）的共聚物显示出增加的转染效率和降低的细胞毒性。(C)1998年Elsevier Science B.V.
Poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) is a water-soluble cationic polymer, which is able to bind to DNA by electrostatic interactions. At a polymer/plasmid ratio above 2 (w/w) positively charged complexes were formed with a size around 0.2 mu m. The transfection efficiency of polymer/plasmid complexes was evaluated in cell culture (COS-7 and OVCAR-3 cells) using a pCMV-lacZ plasmid, encoding for beta-galactosidase, as a reporter gene. The optimal transfection efficiency was found at a PDMAEMA/plasmid ratio of 3-5 (w/w). Under these conditions 3-6% of the cells were actually transfected. Like other cationic polymers, PDMAEMA is slightly cytotoxic. This activity was partially masked by complexing the polymer with DNA. A pronounced effect of the molecular weight of the polymer on the transfection efficiency was observed. An increasing molecular weight resulted in an increasing number of transfected cells. Dynamic light scattering experiments showed that high molecular weight polymers (M-w>300 kDa) were able to condense DNA effectively (particle size 0.15-0.20 mu m). In contrast, when plasmid was incubated with low molecular weight PDMAEMA, large complexes were formed (size 0.5-1.0 mu m). Copolymers of DMAEMA with methyl methacrylate (MMA), ethoxytriethylene glycol methacrylate (triEGMA) or N-vinyl-pyrrolidone (NVP) also acted as transfection agents. A copolymer with 20 mol % of MMA showed a reduced transfection efficiency and a substantial increased cytotoxicity compared with a homopolymer of the same molecular weight. A copolymer with triEGMA (48 mol %) showed both a reduced transfection efficiency and a reduced cytotoxicity, whereas a copolymer with NVP (54 mol %) showed an increased transfection efficiency and a decreased cytotoxicity as compared to a DMAEMA homopolymer. (C) 1998 Elsevier Science B.V.