Abstract: Objective To investigate the feasibility of electroporation for modRNA transfection and to screen the optimal voltage and pulse time required for different cell electroporation. Methods ①The optimal voltage and pulse time parameters for electroporation of EGFP modRNA into human fibroblasts were screened;②Human fibroblasts were transfected with nGFP and EGFP+mCherry modRNA;③Hela, 293T, 3T3, three common adherent cells were electroporated with EGFP modRNA. Results ①The optimum voltage for electroporation of human fibroblasts was 440 V, and the optimal pulse time was 30 ms;②nGFP could be expressed in the nucleus;EGFP and mCherry proteins could be expressed simultaneously in the cytoplasm;③The optimum voltages for Hela, 293T and 3T3 cells were 425 V, 400 V and 440 V respectively, and the optimal pulse time was 30 ms. Conclusion This experiment demonstrates:①The feasibility of transfection of modRNA into cytoplasm and nucleus by electroporation;②Simultaneous introduction of two modRNAs into the same cell and expression;③ Optimal voltage and optimal pulse time are based on different cell types.

Key words: modRNA, Electroporation, Optimal voltage, Optimal pulse time