Study on the Condition of Transfection of Human Ventricular Muscle Ac16 Cells with Large Plasmid
LI Yahuan, FAN Chenghao, XIANG Chenying, LIAN Hong*
The aim of this study was to explore the transfection method and optimum condition of large plasmid transfecting human ventricular muscle Ac16 cell. The plasmid pLenti-CasRx-EGFP was transfected into Ac16 cells by three methods: Lip3000 transfection, Hieff Trans™ Liposomal transfection and lentivirus transfection. The transfection efficiency of exogenous gene was determined by analyzing the proportion of positive cells expressing EGFP. Finally, the optimal transfection method and transfection condition of large plasmid pLenti-CasRx-EGFP was explored. For Ac16 cells, the optimum transfection ratio of plasmid to Lip3000 was 1:2; the optimal transfection ratio of plasmid to Hieff Trans™ Liposomal transfection was 1:3; the optimal MOI value infected with lentivirus was 200. The transfection efficiency of lentivirus was the highest by comparing the efficiency of the three transfection methods. It is concluded that lentivirus is the most suitable method for transfection the large plasmid (pLenti- CasRx-EGFP) larger than 10 Kb into Ac16 cells with MOI of 200.
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