Convenient Doxycycline-Inducible cDNA Expression
- Ultra-low background minimizes leakiness in polyclonal cultures so clonal selection is unnecessary
- Optional fluorescent sensor enables real-time monitoring of induction and repression
- Easy-to-use, complete, all-in-one and two-vector inducible lentiviral expression systems
Cellecta InDOXibleTM Tet-Activated cDNA Lentiviral Expression System enables convenient doxycycline-in- ducible cDNA expression.
Available as an all-in-one lentiviral vector system and as a two-vector inducible system
(1) One-Vector Tet-Activated System: The transactivator that regulates gene induction is on the same construct as the inducible gene of interest. Transduction of the single lentiviral construct provides all the elements needed to tightly regulate inducible cDNA expression.
(2) Two-Vector System: The inducible cDNA that is un- der the control of the responsive promoter is on a vec- tor without the transactivator. The transactivator is on its own vector and transduced separately into cells, and so can be used to create a panel of transactivator cells set up for introduction of different genes of interest.
How Does It Work?
Fusion of a bacterial tet-repressor protein with the VP16 activating domain creates a transactivator that binds, in a tetracycline-dependent manner, the tet-responsive element (TRE) derived from bacterial promoters. Cellecta has optimized the transactivator to ensure very tightly regulated activation of the minimal CMV promoter driving expression of the cDNA of interest. As a result, selection of low background clones is typically unnecessary. Background levels of antibiotic-selected, transduced polyclonal cell cultures is minimal.
The two variations of the tetracycline activator (tTA and rtTA) enable activation with either the addition or removal of the tetracycline analog doxocycline (dox). Since all the elements are provided on lentiviral vectors, they can be easily and effectively introduced into any mammalian cell line.
Fluorescent Sensor for Real-Time Induction Monitoring
A fluorescent reporter (RFP), linked by an internal ribosome entry site (IRES) to the cDNA transcript,
is available with both the single-vector and dual- vector InDOXible Tet-Activated vectors. With this IRES configuration, cell fluorescence increases or decreases in parallel with the cloned cDNA, allowing monitoring of induction. This fluorescent sensor feature allows convenient confirmation of gene modulation during experiments.
For Research Use Only.
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