Hi,I have my gene of interest in pcDNA5/FRT/TO vector. Can the pcDNA5FRT/TO vector be used for transient expression of the gene in a common cell line (different from the Flp-In™ T-REx™ System)? If so, do I need to add doxycycline to induce expression? In this case, can you suggest me the timing and concentration of doxycycline?
Thank you,
Francesca
Hi Francesca,
you can use it for transient transfections in common cell lines. pcDNA5 has modified CMV promoter and will be constitutively active in the absence of Tet repressor.
In the presence of Tet repressor, it binds to TetO2 elements within CMV promoter and blocks the expression of the gene. Dox addition prevents binding of TetR and restore CMV promoter activity.
In your case, no TetR -> full gene expression from pcDNA5.
Good luck!
Eugen
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