Co-transfection and tandem transfection of HEK293A cells for overexpression and RNAi experiments |
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Authors: | Xie Zhen-Li Shao Shu-Li Lv Jian-Wei Wang Chang-He Yuan Cheng-Zhi Zhang Wei-Wei Xu Xing-Jun |
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Institution: | College of Life Science and Engineering, Qiqihar University, Qiqihar, Peoples Republic of China. |
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Abstract: | 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. |
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Keywords: | co‐expression co‐transfection rescue RNA interference tandem transfection |
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