Hello,
I was having difficulty in infecting a cell with a gene of interest. Say, for instance, I was able to introduce a CMV on the cell, but I was not successful in introducing my target gene. I was thinking of trying to introduce the gene thru transient transfection since we have a plasmid for that. Is it possible to perform transient transfection on a previously transduced cell line?
Dear cielo,
I don't see any point why transfection should not work on virus transduced cells. Depending on your cells, there may be some defence reaction going on (TLR activation by the virus). But talking of stbly transduced cell lines, those should not show symptomps of acute infection after time, as they do not produce virus particles. So normally should work.
Dear Cielo,
I agree with Christian you can perform transient transfection. My question is how do you know CMV is inserted but not the gene of interest?
Dear Cielo,
It's better to use the new cell,not the previous ones. But how do you know that your transduction doesn't work for your gene of interest. I remember that my target gene has the GFP marker which I checked it after 48h. Also, it is important to have a control : one sample for transduction with the target gene and the other without it. So, in this case you can say that your target gene has some problems.
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