title: Co-transfection and tandem transfection of HEK293A cells for overexpression and RNAi experiments.
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.
authors.affiliation: College of Life Science and Engineering, Qiqihar University, Qiqihar, Peoples Republic of China.
authors.forename: Zhen-Li
authors.initials: ZL
authors.lastname: Xie
authors.forename: Shu-Li
authors.initials: SL
authors.lastname: Shao
authors.forename: Jian-Wei
authors.initials: JW
authors.lastname: Lv
authors.forename: Chang-He
authors.initials: CH
authors.lastname: Wang
authors.forename: Cheng-Zhi
authors.initials: CZ
authors.lastname: Yuan
authors.forename: Wei-Wei
authors.initials: WW
authors.lastname: Zhang
authors.forename: Xing-Jun
authors.initials: XJ
authors.lastname: Xu
mesh_terms: D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D010957:Plasmids / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins