Hi all,

I have transduced embryonic stem cells with a lentiviral Tet-on system that, upon addition of doxycycline, should express my gene of interest together with GFP. However, when I add dox, I don't see any GFP, nor do I detect an increase in expression of GFP or my GOI by qPCR. After transduction the cells were selected for with puromycin and PCR of the genomic DNA did show that my construct is integrated in the cells.

I have used the construct before in fibroblasts and HEK-293 cells and there it worked without any problems.

Could it be that the tet-one system does not work in certain cell types? And if so, is there a way to solve this?