Theresa Pohlkamp @Theresa-Pohlkamp

In the tet-ON tet-OFF system the target gene is controlled by a TRE (tet responsive element) promoter. It has CMV promoter elements and a few tetO sequences in tandem. Depending on tet-ON (rtTA) or tet-OFF (tTA) the gene of interest is turned on or off with tetracycline/doxycycline, respectievely. My question is, if the TRE-tight promoter, when cloned into the R26 locus, is tight? Or does the Rosa26 region, which acts as a pretty good promoter itself, overrule the TRE-tight? Anybody experience with that? Or is there an alternative safe spot for targeted insertion that people use for the TRE-gene expression? In Sun et al., 1012 Article Inducible and reversible regulation of endogenous gene in mouse

they knocked TRE into the first intron of Nmyc resulting in endogenous Nmyc expression when dox (tet-OFF system) was added and overexpression without dox. I wonder if downstream of a gene/last exon would be a safe spot? Thoughts? Experience?