Retro-X Tet-One inducible expression systems
The Retro-X 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 systems, both the doxycycline-responsive transactivator and your gene of interest are combined onto the same vector.
The Retro-X 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 systems, both the doxycycline-responsive transactivator and your gene of interest are combined on the same vector. This allows for a simpler protocol 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.
Retro-X Tet-One systems are supplied with our Retro-X Universal Packaging System, which produces high titers of MMLV retrovirus and allows you to customize the tropism of your retrovirus to transduce any cell line or primary cell culture efficiently, and thus greatly expands the applications for these inducible 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 of 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 third-generation version of the technology, with the tightest TRE promoter and most sensitive transactivator.
In Tet-On systems, gene expression is activated by the addition of Dox to the growth medium.
Retro-X Q vector technology
The vectors in these systems feature our Retro-X Q-series technology, which:
- Drives high-titer virus production in packaging cells
- Inactivates viral LTRs in target cells to prevent promoter interference
- Single-vector format for a simple protocol
- Outstanding induction control and extremely low basal expression for a broad range of dividing cell types
- TRE3G promoter is optimized for very tight transcriptional control
- Supplied with our best-performing retroviral packaging system (Retro-X Universal)
- Two-vector Tet-On 3G format also available
- Establish a tetracycline-inducible gene expression system in a wide variety of dividing cells using retrovirus
Heinz, N. et al. Retroviral and transposon-based tet-regulated all-in-one vectors with reduced background expression and improved dynamic range. Hum. Gene Ther. 22, 166–76 (2011).
Additional product information
Please see the product’s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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