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UK Health Security Agency

X1/5

X1/5

Catalogue No.

95051229

Cell Line Name

X1/5

Cell Line 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

Characteristics

Tissue of Origin

Cervix

Karyotype

Not specified

Applications

Specific and stringent control of the activity of an individual gene (luciferase) in a mammalian cell can be monitored. It is also suitable for creation of

Disease

Adenocarcinoma

Culture Conditions

Cell Type

Epithelial

Subculture Routine

Split sub-confluent cultures (70-80%) 1:5 to 1:20 i.e. seeding at 5x1,000 to 2x10,000 cells/cm² using 0.05% 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.

Culture 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).

Growth Mode

Adherent

Additional Info

Depositor

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

Country of Origin

Germany

GMO Status

Genetically Modified Organism Class 1 (GMO1)

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

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.

Available Formats

  • Frozen

If use of this culture results in a scientific publication, it should be cited in the publication as: X1/5 (ECACC 95051229).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.