The Tet-One System allows you to generate inducible expression systems as simply as constitutive expression systems. Previous inducible expression systems required transduction of two separate vectors into your cells. With the Tet-One system, both the doxycycline-responsive transactivator and your gene of interest are combined onto the same vector. This allows for greater transduction efficiency, while maintaining the low background expression and up to 25,000-fold induction (Heinz et al., 2011) in response to doxycycline that is found in the Tet-On 3G systems.
How it Works
Doxycycline-controlled “transactivators” are fusion proteins that contain a DNA-binding TetR domain joined to three minimal transcription activation domains from VP16 (Tet-On 3G, Tet-On Advanced and Tet-Off Advanced proteins). With very high specificity, these transactivators recognize and bind to a Tetracycline Response Element (TRE) that is located in the PTRE3GV promoter upstream your gene of interest. Binding of the transactivator to the TRE promoter results in strong activation of the downstream gene. Tet-On 3G is the premium 3rd generation version of the technology, with the tightest TRE promoter and most sensitive transactivator.
In Tet-One systems, gene expression is activated by the addition of Dox to the growth medium.
Ideal Retroviral Packaging
Retro-X Tet-One systems are supplied with our Retro-X Universal Packaging System (Cat. No. 631530) which produces high titers of MMLV retrovirus and allows you to customize the tropism of your retrovirus to efficiently transduce any cell line or primary cell culture, and thus greatly expands the applications for these inducible systems.
Retro-X Q Vector Technology
The vectors in these systems feature Clontech’s Retro-X Q-Series technology, which:
- Drives high titer virus production in packaging cells
- Inactivates viral LTRs in target cells to prevent promoter interference