InvivoGen has developed human cell lines for Tet-inducible protein expression.
RepTor™ cell lines are derived from HEK293 (human embryonic kidney cells) or A549 (human lung carcinoma) cells, two widely used cellular models:
– HEK-RepTor™ cells
– A549-RepTor™ cells
These cells stably and constitutively express an optimized tetracycline repressor (TetR) construct. They are readily transfectable with a pTiGer plasmid, carrying a gene of interest (GOI) cloned downstream of the tet-inducible CMV-EF1-TRE promoter. GOI expression is induced upon addition of doxycycline, a synthetic analog of the tetracycline antibiotic, leading to derepression of the CMV-EF1-TRE promoter. Low doxycycline concentration, ranging from 1 to 10 ng/ml, is sufficient for full induction, thus limiting the antibiotic side effects.
The TiGer Tet-on system is specifically designed for on-demand expression of cytotoxic genes.
It has been engineered to guarantee maximal repression of the GOI, ensuring minimal leakage without doxycycline, and strong GOI expression with doxycycline.
Applications
InvivoGen’s TiGer Tet-on system is useful for, but not restricted to the following applications:
- Controlled expression of cytotoxic genes
- Fine-tuned expression of a GOI
- Screening of toxic protein inhibitors
RepTor™ cells are functionally validated by transient transfection with a pTiGer-reporter plasmid expressing either SEAP or Lucia luciferase.
They are also extensively tested for viability, stability, and absence of mycoplasma to ensure strong and reproducible results. Moreover, we provide detailed handling and experimental procedures to minimize the need for optimization or troubleshooting by the end-user.
