Description: X1/5 is a HeLa cell line stably transfected with a tet repressor fused with the activating domain of virion protein 16 of herpes simplex virus thus generating a tetracycline-controlled transactivator (tTA) This stimulates transcription from a minimal promoter sequence derived from the human cytomegalovirus promoter IE combined with tet operator sequences. Integration of a luciferase gene controlled by a tTA-dependent promoter into this tTA producing HeLa cell line leads to high levels of tetracycline-sensitive luciferase expression. The luciferase activity can be regulated over up to five orders of magnitude depending on the concentration of tetracycline in the culture medium (0-1ug/ml). Specific and stringent control of the activity of an individual gene (luciferase) in the mammalian cell can be monitored. It is also suitable for creation of on/off situations and analysis of mRNA decay rates under physiological conditions

Also Known As:

Species: Human

Tissue: cervix

Growth Properties: Adherent

Morphology: Epithelial

Growth Medium: EMEM (EBSS) + 2mM Glutamine + 1% Non Essential Amino Acids (NEAA) + 1% Vitamins + 500µg/ml G418 + 300µg/ml Hygromycin + 10% Foetal Bovine Serum (FBS). Freeze cells in 40% medium, 10% Dimethyl Sulphoxide (DMSO) and 50% Foetal Bovine Serum (FBS).

Subculturing Procedure: Split sub-confluent cultures (70-80%) 1:5 to 1:20 i.e. seeding at 5×1,000 to 2×10,000 cells/cm² using 0.25% trypsin or trypsin/EDTA. Alternatively cells can be washed with PBS and subcultured after addition of PBS with 0.8mM EDTA (30% of original volume); 5% CO2; 37°C.

Release Conditions: No

Depositor: Dr M Gossen and Prof Dr H Bujard, Zentrum für Moleculaire Biologie, University Heidelberg

Originator: Yes

References: Gossen M, Bujard H. 1992 Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A. 89(12):5547-51 PMID: 1319065.

Hyperlink to ECACC Cell Line Data Sheet: 95051229