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Inducible Vectors FAQs
1. How does the Tet-On 3G Inducible System work?
OriGene's All-in-one Tet-On system is a new and improved version of the original Tet-On systems designed to significantly stimulate expression of the downstream gene of interest (GOI). It has a Tet-On 3G transactivator and a tightly regulated TRE promoter (PTIGHT) in one vector. The Tet-On 3G transactivator consists of a modified bacterial Tet repressor (TetR) fused to three minimal VP16 activation domains to create a transcriptional activator protein. Our Tet-On 3G transactivator contains mutations that significantly increase its sensitivity to Doxycycline (Dox), a synthetic analog to tetracycline. The increased sensitivity is particularly advantageous for in vivo studies in tissues where high Dox concentrations are difficult to attain (e.g., brain).
2. Can I use tetracycline in the Tet-On system?
Yes, but doxycycline is more stable in the culture medium (48 hours versus 24 hours for tetracycline) and appears to be more potent than tetracycline.
3. What are the applications of Tet-On vectors?
Our Tet-ON system can be used to investigate the role of your gene of interest with relation to progression of a disease, cell cycle, or tissue growth. Virtually any experiment requiring regulation of gene expression is easily accomplished with the use of our new and improved Tet-ON system.
4. Do you have Lentiviral Tet-On vectors?
Yes! Our Lentiviral Tet-On vectors are exclusively available through our custom lentiviral packaging service, so you’ll receive transduction-ready viral particles instead of the vectors themselves.
5. How do we test for leaking in the Tet-on vectors?
We have confirmed that there is no leaking issue within our Tet-On vectors by Western blotting analysis and GFP imaging as we didn’t observe any expression of target proteins without doxycycline.
6. Can you make inducible cell lines using Tet-On vectors?
Yes, our Tet-On vectors could be used for generation of stable cell lines. You can either make them yourself or go through our popular stable cell line development service!