Optimization of Liposome-mediated Transfection of CT26 Cells Based on Functional Identification of Pi-deficiency Syndrome Relative Gene RPS20

Objective To investigate the optimum transfection conditions of cationic liposome transfected CT26 cells(murine colon cancer cells),thus to provide reference for functional identification of spleen deficiency-related gene RPS20 using RNA interfering methods in mice.Methods We transfected pcDNATM6.2-GW/EmGFP-miR-neg Control plasmid into CT26 cells which planted in six-well plate with cationic liposome,and observed the effect of plasmid and Lipofectamine2000 in various proportions on transfection efficiency by detecting fluorescence expression using fluorescence microscopy and flow cytometry.Results The results of detecting transfection efficiency by fluorescence microscope showed that plasmid 4.0μg and 6.0μg had no significant difference in transfection efficiency.The results of flow cytometry showed that the highest transfect efficiency(average being 25.3%)was obtained in the group of Lipofectamine2000 15.0μL+plasmid 4.0μg.Conclusion The optimum transfection condition is tranfection into CT26 cells with plasmid 4.0μg+Lipofectamine2000 15.0μL in six-well plate.