Effect of size and serum proteins on transfection efficiency of poly((2-dimethylamino)ethyl methacrylate)-plasmid nanoparticles
Effect of size and serum proteins on transfection efficiency of poly((2-dimethylamino)ethyl methacrylate)-plasmid nanoparticles
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DOI：
10.1023/a:1016054623543
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发表时间：
1996-07-01
影响因子：
3.7
通讯作者：
Hennink, WE
中科院分区：
文献类型：
--
作者：
Cherng, JY;vandeWetering, P;Hennink, WE
目的。本研究的目的是了解由质粒和合成的阳离子聚合物(聚(2-二甲氨基)甲基丙烯酸乙酯，PDMAEMA)形成的颗粒的物理特性与它们的转染率之间的关系。用动态光散射(大小)和凝胶迁移率测量(电荷)对PDMAEMA-质粒进行了表征。以编码β-半乳糖苷酶的pCMV-LacZ为报告基因，在COS-7细胞中检测其转染率。结果表明，当PDMAEMA-质粒比为3(w/w)时，转染率最高，得到的颗粒较均匀，粒径为0.15微米，粒径分布较窄，且带有少量正电荷。DLS测量表明，在较低重量比下制备的颗粒显示出较低的转染率，并且在时间上不稳定。与其他阳离子聚合物一样，PDMAEMA具有轻微的细胞毒性。通过将聚合物与DNA络合，这一活性被部分掩盖。有趣的是，该颗粒的转染率不受血清蛋白存在的影响。PDMAEMA是一种有趣的载体，可用于设计体内和体外的基因转染系统。
Purpose. The aim of this study was to gain insight into the relation between the physical characteristics of particles formed by a plasmid and a synthetic cationic polymer (poly(2-dimethylamino)ethyl methacrylate, PDMAEMA) and their transfection efficiency.Methods. The PDMAEMA-plasmid particles were characterized by dynamic light scattering (size) and electrophoretic mobility measurements (charge). The transfection efficiency was evaluated in cell culture (COS-7 cells) using a pCMV-lacZ plasmid coding for beta-galactosidase as a reporter gene.Results. It was shown that the optimal transfection efficiency was found at a PDMAEMA-plasmid ratio of 3 (w/w), yielding stable and rather homogeneous particles (diameter 0.15 mu m) with a narrow size distribution and a slightly positive charge. Particles prepared at lower weight ratios, showed a reduced transfection efficiency and were unstable in time as demonstrated by DLS measurements. Like other cationic polymers, PDMAEMA is slightly cytotoxic. This activity was partially masked by complexing the polymer with DNA. Interestingly, the transfection efficiency of the particles was not affected by the presence of serum proteins.Conclusions. PDMAEMA is an interesting vector for the design of in vivo and ex vivo gene transfection systems.