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Co‐transfection and tandem transfection of HEK293A cells for overexpression and RNAi experiments
Author(s) -
Xie ZhenLi,
Shao ShuLi,
Lv JianWei,
Wang ChangHe,
Yuan ChengZhi,
Zhang WeiWei,
Xu XingJun
Publication year - 2011
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1042/cbi20100470
Subject(s) - transfection , rna interference , microbiology and biotechnology , tandem , chemistry , biology , gene , biochemistry , rna , materials science , composite material
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.