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Are there any issues with tetracycline inducible promoters popping on all of a s


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#1 Ahrenhase

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Posted 10 February 2014 - 12:36 PM

I'm designing a transgene that contains cre recombinase under the control of a tet inducible promoter.  This entire transgene will be flanked by lox P sites.  Therefore, this transgene will negative regulate itself.  I'm just concerned about this promoter getting turned by other factors.  I know tetracycline is found in some FBS, so I will need to use verified tet-free FBS.  Are there any other caveats to this method?



#2 bob1

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Posted 10 February 2014 - 01:59 PM

I've found tet inducible promoters to be a bit leaky in mammalian cell culture, so even in the absence of tet you are likely to get a low level expression.



#3 Ahrenhase

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Posted 10 February 2014 - 06:31 PM

I've found tet inducible promoters to be a bit leaky in mammalian cell culture, so even in the absence of tet you are likely to get a low level expression.

I know there are DEX/DOX systems that allow for tighter control.  I wanted to avoid DEX because of off-target effects it may have.  DOX has off target effects too, but I figured my cell cultures are seeing DOX in the media (which contains FBS) anyway. 






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