What promoter should be used to express Tet-On 3G transactivator? CMV or EF1 alpha promoter?

Tet-inducible expression in stem cells and hematopoietic cells: EF-1 alpha systems Some types of cells (particularly hematopoietic cells and stem cells) tend to silence CMV promoters over time. EF-1 alpha promoter versions of the Tet-On 3G systems provide consistent long-term expression of the Tet-On 3G transactivator in these cells, as well as a broad range of other cell types The Tet-On 3G [...]

What are the features of the Tet-On 3G transcriptional activator: highest sensitivity to doxycycline

Tet-On 3G is a modified, improved Tet-On Advanced transcriptional activator protein (transactivator) which has been evolved to display far higher sensitivity to doxycycline. Tet-On 3G generates very high maximal expression and responds to lower Dox concentrations than its predecessors. These Dox concentrations are far below cytotoxic levels for either cell culture or transgenic studies. The [...]

What is the Tet-On 3G transactivator?

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 (transactivator). Five amino acid changes convert Takara Bio's Tet-On Advanced transactivator to Tet-On 3G. Tetracycline-inducible expression systems Care guide Caution [body] This care sheet provides general information [...]

What are the features of the pTRE3G inducible promoter: tetracycline-inducible expression, lowest basal activity level?

The tetracycline responsive element (TRE) consists of seven repeats of a 19-bp tet operator sequence located upstream of a minimal CMV promoter. It provides very low basal expression and high maximal expression after induction. Although the tet operator repeats are identical in all Tet-On generations, the junction sequences in the Tet-On 3G version (PTRE3G) have been altered to an even spacing [...]

What are the advantages of Takara Tet-On 3G systems?

When the Tet-On system was originally launched by Clontech Laboratories (now Takara Bio USA, Inc.) in 1996, it was the premier inducible expression system. However, its performance has been surpassed by two subsequent generations of the Tet-On system. Compared to their predecessors, Tet-On 3G systems demonstrate lower basal expression and higher sensitivity to the doxycycline (Dox) inducer. 1. [...]