Gene Expression of Aspect Ratio-Controlled Polyplexes Based on the Effect of Multi-Arm Poly(ethylene glycol)
Gene Expression of Aspect Ratio-Controlled Polyplexes Based on the Effect of Multi-Arm Poly(ethylene glycol)
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DOI：
10.1021/acsbiomaterials.8b01498
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发表时间：
2019-11-01
影响因子：
5.8
通讯作者：
Kono, Kenji
中科院分区：
文献类型：
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作者：
Harada, Atsushi;Nomura, Kenta;Kono, Kenji
通过将pDNA与聚（L-赖氨酸）（PLL）均聚物和末端带有多臂聚（乙二醇）（PEG）部分的PLL（mapPEG-PLL）的聚阳离子混合物混合制备的复合物的纵横比的控制被检查。通过改变聚阳离子混合物中的mapPEG-PLL含量，分别通过染料排阻测定和AFM观察来评估伴随着聚合物形成的pDNA的缩合和聚合物的形态。增加maPEG-PLL含量引起聚合物的伸长，并且通过控制maPEG-PLL含量成功地制备了纵横比为2至10的聚合物。通过琼脂糖凝胶电泳和实时PCR测量，分别检查pDNA在聚合复合物中具有不同的纵横比对DNA酶I和聚合酶的反应性。此外，通过HeLa细胞的聚合复合物的细胞摄取和转染效率进行了评估。结果表明，聚合复合物的纵横比的增加引起PCR效率的增加，伴随着细胞摄取的减少。
Controlling the aspect ratio of polyplexes prepared by mixing pDNA with a polycation mixture of a poly(L-lysine) (PLL) homopolymer and PLL terminally bearing a multiarm poly(ethylene glycol) (PEG) part (maPEG-PLL) was examined. By varying the maPEG-PLL content in the polycation mixture, the condensation of pDNA accompanied by polyplex formation and the morphology of the polyplexes were evaluated by a dye exclusion assay and AFM observations, respectively. Increasing the maPEG-PLL content caused elongation of the polyplex, and polyplexes with aspect ratios from 2 to 10 were prepared successfully by controlling the maPEG-PLL content. The reactivity of pDNA in the polyplexes with varying aspect ratios against DNase I and polymerase were examined by agarose gel electrophoresis and real-time PCR measurements, respectively. Moreover, cellular uptake and transfection efficiency of the polyplexes by HeLa cells was evaluated. The results revealed that an increase in aspect ratio of the polyplexes caused an increase in PCR efficiency with a concomitant decrease in cellular uptake.