Since the publication of the tTA- (1) and more recently the rtTA-based (2) expression system, many researchers have sucessfully made use of the two systems to control the activity of their gene of interest with tetracycline in cell cultures as well as in transgenic organisms. We nevertheless fielded a number of inquiries in which often the same questions were asked. In response to this, we have prepared a "troubleshooting guide". We have also compiled once more all relevant information on the plasmid family which is required to establish the system. Both will hopefully be useful for you and help you avoid some of the pitfalls and problems one may encounter. For some more general considerations, we refer you to ref. 3.
During the last two years we have sent out more than 5000 probes of plasmids and many samples of cell lines. This has been quite a burden to our lab. Thus, please understand that we cannot in addition answer questions concerning specifics of your projects, in particular since we often realize that inquiries often concern issues we have dealt with in our publications. For the same reasons please also refrain from calling us up. In our publications and in the following you will find all the information required for setting up the tet systems in tissue culture. In the forthcoming update we will expand this guide into the area of mouse transgenics.
Good luck in your experiments.
The Bujard group