Comparing Chemical Transfection, Electroporation, and Lentiviral Vector Transduction to Achieve Optimal Transfection Conditions in the Vero Cell Line

Abstract

Abstract Background: Transfection is an important analytical method for studying gene expression in the cellular environment. There are some barriers to efficient DNA transfection in host cells, including circumventing the plasma membrane, escaping endosomal compartmentalization, autophagy, immune sensing pathways, and translocating the nuclear envelope. Therefore, it would be very useful to introduce an optimum transfection approach to achieve high transfection efficiency in the Vero cell line.The aim of this study was to compare various transfection techniques and introduce a highly efficient method for gene delivery in the Vero cell line. Methods: In the current study, three transfection methods were performed, including chemical transfection, electroporation, and lentiviral vector transduction, to obtain the optimum transfection conditions in the Vero cell line. Vero cells were cultured and transfected with chemical transfection reagents, electroporation, and HIV-1-based lentivectors under different experimental conditions. Transfection efficiency was assessed using flow cytometry and fluorescence microscopy to detect GFP-positive cells. Results: Among the tested methods, TurboFect™ chemical transfection exhibited the highest efficiency. Optimal transfection conditions were achieved using 1 μg DNA and 4 µL TurboFect™ on 6×104 Vero cells. Conclusion: TurboFect™, a cationic polymer transfection reagent, demonstrated superior transfection efficiency in Vero cells compared with electroporation and lentivirus particles and is the optimal choice for chemical transfection in the Vero cell line.