Abstract:

Abstract: At the core of gene therapy lies the use of viral vectors, engineered viruses serving as delivery vehicles to transport restorative genes into target cells. Therefore, the effect of 7 different rAAV serotypes and their different quantites was analysis here on human prostate cancer cell lines PC-3 and DU-145, which are hard to be transfected. PC-3 and DU-145 cell lines were infected with different multiplicity of infection (MOI) ratios of 7 rAAV serotypes, AAV 2/1, 2/2, 2/3, 2/5, 2/6, and 2/9, which were expressing the green fluorescent protein (GFP) transgene driven by the CMV promoter. The transduction efficiency was analyzed by fluorescent microscopy and flow cytometry. In addition, the cell viability of the infected cells was measured by Muse Cell Analyzer at the MOI of 10.000. rAAV 2/2 and rAAV 2/6 have the most significant ability to transduce PC-3 cells. Although rAAV 2/2 and rAAV 2/6 were also the most transducing serotypes in the DU-145 cell line, the transduction rates did not exceed 20% in this cell line. On the other hand, after viral infection, no difference in cell viability was observed in PC-3 cells compared to the mock group, while a significant decrease in viability was observed in DU-145 cells. This study determined the transduction efficiency of 7 different rAAV serotypes on human cancer cell lines. While rAAV 2/2 and rAAV 2/6 serotypes achieved more than 60% transduction efficiency in PC-3 cells, the transduction efficiency could not exceed 20% in DU-145 cells. Overall, this study demonstrated that rAAV 2/2 and rAAV 2/6 could mediate the expression of a transgene with a high transduction efficiency.

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