I have been trying to express a gene (GFP) in a retroviral system by transfecting human Phoenix-ECO cells, and transducing murine PG13 cells with the supernatant. I then sort GFP-positive PG13 cells, and culture them. Once I use harvested viral sup from sorted PG13 cells (after 12-24 hours) to transduce several different human cell lines, I see little or no GFP expression in the days after transduction. I know that temperature may have a role in viral expression (37 vs 32 degrees), but this cannot be the only factor leading to poor efficiency. What are other ways to improve transduction efficiency?
I centrifuged the supernatant, used the concentrated sample for transduction, and got good GFP and protein expression in cell culture and animal models. However, I used rat and mice cells.
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