College of Life Science and Engineering, Qiqihar University, Qiqihar, Peoples Republic of China.
Cell Biol Int. 2011 Mar;35(3):187-92. doi: 10.1042/CBI20100470.
pIRES2-EGFP was employed and a non-target shRNA expressing plasmid was constructed to simulate overexpression and RNAi (RNA interference) experiments. Transfection of pIRES2-EGFP into HEK293A cells by cationic lipids VigoFect demonstrated that transfection efficiency increased in a dose-dependent manner with amount of DNA plasmid used, and optimal transfection time and cell density should be identified to reach a compromise of higher transfection efficiency and lower toxicity. Co-transfection experiments indicated that the two co-transfected plasmids were equivalently delivered into the same cells, and the co-transfection efficiency was rarely affected by cell density and proportion of the two plasmids. However, plasmid-receipted cells seemed indisposed to accept plasmid again during the second transfection, and very low co-transfection efficiency was observed in tandem transfection.
pIRES2-EGFP 被采用，构建了一个非靶向 shRNA 表达质粒来模拟过表达和 RNAi（RNA 干扰）实验。阳离子脂质体 VigoFect 将 pIRES2-EGFP 转染到 HEK293A 细胞中，证明转染效率随着使用的 DNA 质粒数量的增加呈剂量依赖性增加，并且应该确定最佳的转染时间和细胞密度，以达到更高的转染效率和更低的毒性之间的平衡。共转染实验表明，两种共转染的质粒被等量递送到相同的细胞中，并且共转染效率很少受到细胞密度和两种质粒比例的影响。然而，质粒接受的细胞似乎在第二次转染时不愿意再次接受质粒，并且在串联转染中观察到非常低的共转染效率。
