In article <46nifu$3f5 at mserv1.dl.ac.uk>, grggta at picr.cr.man.ac.uk says...
[SNIP - to save space]
[Repression in tet transactivator system is cell-type dependent]
[Failure to generate stable cell lines based on this system]
>Please anyone let me know if the have similar problems, any solutions
>with this system.
Since the authors managed to generate transgenic animals based on this
system I assume that stable cells are a possibility
Furth et al. PNAS 91, 9302-9306 (1994)
I think maybe what you are after is the modified system described recently
Shockett et al. PNAS 92, 6522-6526 (1995)
The authors show that the tTA transactivator is toxic so they put the
expression under control of an autoregulatory loop. Although not
perfect it seems to be a better bet than the original when it comes
to cell lines and transgenic mice.
On a similar note, is anyone trying mouse lines based on the Lac Switch
system (Stratagene)? I was hoping that someone might generate mice with
promiscuous LacI expression so that they could be used to generate
switched mouse lines upon introduction of the appropriate constructs.
I realise that the in vivo pharmacokinetics of IPTG are not the best
(half life of 30 min) but it should be ideal for transient induction.
Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)