|Cell Line Name:||U5A|
|Keywords:||alpha interferon signalling pathway, IFN regulated human gene 6-16,|
|Cell Line Description:||The human sarcoma cell line HT 1080 (ECACC Catalogue number 85111505) was transfected with a vector encoding a selectable marker (guanine phosphoribosyltransferase) regulated by alpha interferon to create the 2fTGH cell line, enabling the selection of mutations in genes encoding components of the interferon signalling pathway. Chemical mutagenesis of the parent 2fTGH cell line enabled the isolation of a number of IFNg signalling mutants available from ECACC: U2A (12021502), U3A (12021503), U4A (12021504), U4C (12021505), U5A (12021506), U6A (12021507). Each contains a different mutation in the IFN signalling pathway. The 2fTGH panel provides a tool for the in vitro study and comparison of disrupted interferon signalling at multiple points across the IFN pathway. The U5A mutant cell line lacks the ifnar2-2 component of the interferon-alpha and interferon-beta receptor and the beta-R1 gene product. U5A is completely defective in interferon alphabeta binding and response.|
|GMO Status:||Genetically Modified Organism Class 1 (GMO1)|
|Subculture Routine:||Split sub-confluent cultures (70-80%) 1:4 to 1:8 i.e. seeding at 1-4x10,000 cells/cm² using 0.25% trypsin/EDTA; 5% CO2; 37°C.|
|Culture Medium:||DMEM + 2mM Glutamine + 10% Foetal Bovine Serum (FBS).|
|Depositor:||Licensed from: Cancer Research Technology Ltd Angel Building 407 St John Street London EC1V 4AD. Originator: Dr GR Stark, Imperial Cancer Research Fund, Lincoln's Inn Fields, London WC2A 3PX|
|References:||References on transfection: Blochlinger K, Diggelmann H.1984 Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells. Mol Cell Biol. 4(12):2929-31. PMID: 6098829. Kelly JM, Porter AC, Chernajovsky Y, Gilbert CS, Stark GR, Kerr IM.1986 Characterization of a human gene inducible by alpha- and beta-interferons and its expression in mouse cells. EMBO J. 5(7):1601-6. PMID: 3017706. Porter AC, Chernajovsky Y, Dale TC, Gilbert CS, Stark GR, Kerr IM.1988 Interferon response element of the human gene 6-16. EMBO J. 7(1):85-92. PMID: 3359997.|
|Additional Bibliography:||Lutfalla G, Holland SJ, Cinato E, Monneron D, Reboul J, Rogers NC, Smith JM, Stark GR, Gardiner K, Mogensen KE, et al. 1995 Mutant U5A cells are complemented by an interferon-alpha beta receptor subunit generated by alternative processing of a new member of a cytokine receptor gene cluster. EMBO J. 14(20):5100-8. PMID: 7588638.|
|Patents:||None specified by Depositor|
|Research Council Deposit:||No|
|Release Conditions:||Yes - commercial organisations are required to complete the 'Cell Line Release Authorisation for Research Use in Commercial Organisations' release conditions form.|
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