Recently, we constructed a transgenic mouse model, in which the transgenic microRNA expression is controlled by the tet on system. So we added doxycycline to the drinking water, but i am not sure what the optimal time used for inducing microRNA expression is and I don't know when the time to add doxycycline to the water is. Is it ok to start it when the mouse is 8 weeks old? And the most important thing is whether the drug doxycycline could induce the mature microRNA when we detect the mouse by tagman microarray(which just detects the mature microRNA in the tissue). These things really confused me in the experiment, could anyone who is familiar with it give me some suggestions?
Hi Allen,
I think to check the sensitivity of doxycycline you should be known of exact time of expression of miRNA. That could help you to add the doxycycline, if you know the optical time then it absolutely depend on the sensitivity of doxycycline on your transgenic mouse.
Hi Allen, induction of the system by adding Dox is quite rapid, depending on the tissue of interest it reacts in minutes or hours. We could observe a specific Ptet-induced luc signal already after less than 1 minute with an old fashioned imaging system. However maximal induction is only reached after several hours (liver: about 8 hours).
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