Poly(3-guanidinopropyl methacrylate): A novel cationic polymer for gene delivery
Poly(3-guanidinopropyl methacrylate): A novel cationic polymer for gene delivery
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DOI：
10.1021/bc049864q
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发表时间：
2004-11-01
影响因子：
4.7
通讯作者：
Hennink, WE
中科院分区：
文献类型：
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作者：
Funhoff, AM;van Nostrum, CF;Hennink, WE
设计了带有胍侧基的阳离子聚甲基丙烯酸酯，以制造具有细胞膜穿透特性的聚合物，例如 Tat 或其他富含精氨酸的肽。采用自由基聚合法合成了聚合物聚(3-胍基丙基甲基丙烯酸酯)，简称pGuaMA。通过动态光散射和 zeta 电位测量研究了 pGuaMA（分子量 180 kDa）的 DNA 凝聚特性，发现了带正电的小颗粒（110 nm，+37 mV）。结果表明，基于 pGuaMA 的聚合复合物能够在没有血清的情况下有效转染 COS-7 细胞，而在相同条件下，聚（精氨酸）（pArg）聚合复合物没有显示出可检测的转染水平。将源自流感病毒的膜破坏肽 INF 7 添加到预先形成的 pGuaMA 复合物中确实导致转染水平增加了 2 倍。 DLS、zeta 电位测量、凝胶电泳和溴化乙锭位移测量表明，血清在高聚合物/质粒比例下诱导聚合复合物聚集，而在低聚合物/质粒比例下，聚合复合物的极性可能由于其表面吸附带负电的蛋白质而发生逆转。很可能，pGuaMA 复合物与血清蛋白的不利相互作用是这些复合物在血清存在下缺乏转染活性的原因。共焦激光扫描显微镜表明细胞通过聚合复合物和游离聚合物的内吞作用进行内化。因此，正如其他聚合复合物所报道的那样，pGuaMA聚合复合物通过内吞作用进入细胞，而不是如假设的那样通过直接膜通道进入细胞。
A cationic polymethacrylate with a guanidinium side group was designed in order to create a polymer with cell membrane-penetrating properties such as Tat or other arginine-rich peptides. The polymer, poly(3-guanidinopropyl methacrylate), abbreviated as pGuaMA, was synthesized by free radical polymerization. The DNA-condensing properties of pGuaMA (Mw 180 kDa) were investigated via dynamic light scattering and zeta potential measurements, and small, positively charged particles (110 nm, +37 mV) were found. It was shown that polyplexes based on pGuaMA were able to transfect COS-7 cells efficiently in the absence of serum, while under the same conditions poly(arginine) (pArg) polyplexes did not show detectable transfection levels. Addition of a membrane-disrupting peptide, INF 7, derived from the influenza virus, to preformed pGuaMA polyplexes did result in similar to2 times increased transfection levels. DLS, zeta potential measurements, gel electrophoresis, and ethidium bromide displacement measurements indicated that serum induced aggregation of the polyplexes at high polymer/plasmid ratios, while at low polymer/plasmid ratios the polarity of the polyplexes reversed likely due to adsorption of negatively charged proteins on their surface. Likely, the unfavorable interactions of pGuaMA polyplexes with serum proteins is the reason for the absent transfection activity of these polyplexes in the presence of serum. Confocal laser scanning microscopy indicated cellular internalization via endocytosis of both polyplexes and free polymer. Thus, pGuaMA polyplexes enter cells, as reported for other polyplexes, by endocytosis and not, as hypothesized, via direct membrane passage.