What promoter should be used to express Tet-On 3G transactivator? CMV or EF1 alpha promoter?
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
![Express Tet-On 3G from EF1a in stem cells](https://www.takarabio.com/images/360-Tet/CC-TechFigs/TET_ON_3Gexpress_T.jpg)
The Tet-On 3G Inducible Expression System (EF1alpha Version) provides inducible expression in stem cells. Mouse embryonic stem cells were cotransfected with pTRE3G-ZsGreen1 and pEF1a-Tet3G using Xfect mESC Transfection Reagent. The stem cells show ZsGreen1 expression only in the presence of Dox.
![Express Tet-On 3G from EF1a in Jurkat cells](https://www.takarabio.com/images/360-Tet/CC-TechFigs/TET_ON_3Gexpresschart_T.jpg)
The Tet-On 3G Inducible Expression System (EF1alpha Version) provides inducible expression in hematopoietic cells. Jurkat cells were transfected with pEF1a-Tet3G using Xfect Transfection Reagent, and stable clones were selected by limiting dilution. 83% of the Jurkat Tet-On 3G clones that we tested showed strong inducible expression. Such levels of control were not possible with previous versions of Tet-On systems for this cell line.