Research on Transfection Condition of RNA Interference of Intestinal Epithelial Cells Based on Functional Identification of Spleen-deficiency Syndrome Associated Genes

Objective To investigate the transfection condition for affecting RNA interference efficiency on rat intestinal epithelial cells(IEC-6) cells,and to lay a foundation for the further research of constructing cell model of target protein different expression by the method of RNA interference.Methods IEC-6 were cultured in 24-well plate.With the help of cation liposome,the transfection of FAM-siRNA fragment into ICE-6 was carried out,and the effect of FAM-siRNA and Lipofectamine 2000 in different proportions on the transfection efficiency was examined with fluorescence microscope and flow cytometry.Results(1)The results of scoring of transfection efficiency under fluorescence microscope showed that the score was zero when Lipofectamine 2000 or FAM-siRNA was used alone,or both of them missed,but there was weak or strong fluorescence expression when Lipofectamine 2000 and FAM-siRNA used together.(2) The results of detecting transfection efficiency by flow cytometry showed that the transfect efficiency was the highest(76.3% on average) in the group of Lipofectamine 2000 1.0 μL+FAM-siRNA 160 nmol/L,while was only 1% in the blank group.Conclusion Under the transfection condition of FAM-siRNA 160 nmol/L(20 μmol/L,4.8 μL)+ Lipofectamine 2000 1.0 μL,IEC-6 has the highest transfection efficiency during RNA interference.