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Proc Natl Acad Sci U S A. Jun 15, 1991; 88(12): 5096–5100.

Direct derivation of conditionally immortal cell lines from an H-2Kb-tsA58 transgenic mouse.


Studies on cell lines have greatly improved our understanding of many important biological questions. Generation of cell lines is facilitated by the introduction of immortalizing oncogenes into cell types of interest. One gene known to immortalize many different cell types in vitro encodes the simian virus 40 (SV40) large tumor (T) antigen (TAg). To circumvent the need for gene insertion in vitro to generate cell lines, we created transgenic mice harboring the SV40 TAg gene. Since previous studies have shown that TAg expression in transgenic mice is associated with tumorigenesis and aberrant development, we utilized a thermolabile TAg [from a SV40 strain, tsA58, temperature sensitive (ts) for transformation] to reduce the levels of functional TAg present in vivo. To direct expression to a broad range of tissues, we used the mouse major histocompatibility complex H-2Kb promoter, which is both widely active and can be further induced by interferons. tsA58 TAg mRNA was expressed in tissues of all animals harboring the hybrid construct. Development of all tissues was macroscopically normal except for thymus, which consistently showed hyperplasia. Fibroblast and cytokeratin+ thymic epithelial cultures from these mice were readily established without undergoing crisis and were conditionally immortal in their growth; the degree of conditionality was correlated with the levels of tsA58 TAg detected. One strain of H-2Kb-tsA58 mice has been bred through several generations to homozygosity and transmits a functional copy of the transgene.

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  • Jat PS, Sharp PA. Large T antigens of simian virus 40 and polyomavirus efficiently establish primary fibroblasts. J Virol. 1986 Sep;59(3):746–750. [PMC free article] [PubMed]
  • Frederiksen K, Jat PS, Valtz N, Levy D, McKay R. Immortalization of precursor cells from the mammalian CNS. Neuron. 1988 Aug;1(6):439–448. [PubMed]
  • Bayley SA, Stones AJ, Smith CG. Immortalization of rat keratinocytes by transfection with polyomavirus large T gene. Exp Cell Res. 1988 Jul;177(1):232–236. [PubMed]
  • Birren SJ, Anderson DJ. A v-myc-immortalized sympathoadrenal progenitor cell line in which neuronal differentiation is initiated by FGF but not NGF. Neuron. 1990 Feb;4(2):189–201. [PubMed]
  • Burns JS, Lemoine L, Lemoine NR, Williams ED, Wynford-Thomas D. Thyroid epithelial cell transformation by a retroviral vector expressing SV40 large T. Br J Cancer. 1989 May;59(5):755–760. [PMC free article] [PubMed]
  • Evrard C, Galiana E, Rouget P. Immortalization of bipotential glial progenitors and generation of permanent "blue" cell lines. J Neurosci Res. 1988 Sep;21(1):80–87. [PubMed]
  • Lemoine NR, Mayall ES, Jones T, Sheer D, McDermid S, Kendall-Taylor P, Wynford-Thomas D. Characterisation of human thyroid epithelial cells immortalised in vitro by simian virus 40 DNA transfection. Br J Cancer. 1989 Dec;60(6):897–903. [PMC free article] [PubMed]
  • Santerre RF, Cook RA, Crisel RM, Sharp JD, Schmidt RJ, Williams DC, Wilson CP. Insulin synthesis in a clonal cell line of simian virus 40-transformed hamster pancreatic beta cells. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4339–4343. [PMC free article] [PubMed]
  • Williams DA, Rosenblatt MF, Beier DR, Cone RD. Generation of murine stromal cell lines supporting hematopoietic stem cell proliferation by use of recombinant retrovirus vectors encoding simian virus 40 large T antigen. Mol Cell Biol. 1988 Sep;8(9):3864–3871. [PMC free article] [PubMed]
  • Ridley AJ, Paterson HF, Noble M, Land H. Ras-mediated cell cycle arrest is altered by nuclear oncogenes to induce Schwann cell transformation. EMBO J. 1988 Jun;7(6):1635–1645. [PMC free article] [PubMed]
  • Tegtmeyer P. Function of simian virus 40 gene A in transforming infection. J Virol. 1975 Mar;15(3):613–618. [PMC free article] [PubMed]
  • Petit CA, Gardes M, Feunteun J. Immortalization of rodent embryo fibroblasts by SV40 is maintained by the A gene. Virology. 1983 May;127(1):74–82. [PubMed]
  • Jat PS, Sharp PA. Cell lines established by a temperature-sensitive simian virus 40 large-T-antigen gene are growth restricted at the nonpermissive temperature. Mol Cell Biol. 1989 Apr;9(4):1672–1681. [PMC free article] [PubMed]
  • Zaret KS, DiPersio CM, Jackson DA, Montigny WJ, Weinstat DL. Conditional enhancement of liver-specific gene transcription. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9076–9080. [PMC free article] [PubMed]
  • Radna RL, Caton Y, Jha KK, Kaplan P, Li G, Traganos F, Ozer HL. Growth of immortal simian virus 40 tsA-transformed human fibroblasts is temperature dependent. Mol Cell Biol. 1989 Jul;9(7):3093–3096. [PMC free article] [PubMed]
  • Weiss EH, Mellor A, Golden L, Fahrner K, Simpson E, Hurst J, Flavell RA. The structure of a mutant H-2 gene suggests that the generation of polymorphism in H-2 genes may occur by gene conversion-like events. Nature. 1983 Feb 24;301(5902):671–674. [PubMed]
  • Baldwin AS, Jr, Sharp PA. Binding of a nuclear factor to a regulatory sequence in the promoter of the mouse H-2Kb class I major histocompatibility gene. Mol Cell Biol. 1987 Jan;7(1):305–313. [PMC free article] [PubMed]
  • Kimura A, Israël A, Le Bail O, Kourilsky P. Detailed analysis of the mouse H-2Kb promoter: enhancer-like sequences and their role in the regulation of class I gene expression. Cell. 1986 Jan 31;44(2):261–272. [PubMed]
  • David-Watine B, Israël A, Kourilsky P. The regulation and expression of MHC class I genes. Immunol Today. 1990 Aug;11(8):286–292. [PubMed]
  • Wallach D, Fellous M, Revel M. Preferential effect of gamma interferon on the synthesis of HLA antigens and their mRNAs in human cells. Nature. 1982 Oct 28;299(5886):833–836. [PubMed]
  • Israel A, Kimura A, Fournier A, Fellous M, Kourilsky P. Interferon response sequence potentiates activity of an enhancer in the promoter region of a mouse H-2 gene. Nature. 1986 Aug 21;322(6081):743–746. [PubMed]
  • Kollias G, Spanopoulou E, Grosveld F, Ritter M, Beech J, Morris R. Differential regulation of a Thy-1 gene in transgenic mice. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1492–1496. [PMC free article] [PubMed]
  • Noble M, Fok-Seang J, Cohen J. Glia are a unique substrate for the in vitro growth of central nervous system neurons. J Neurosci. 1984 Jul;4(7):1892–1903. [PubMed]
  • Efrat S, Linde S, Kofod H, Spector D, Delannoy M, Grant S, Hanahan D, Baekkeskov S. Beta-cell lines derived from transgenic mice expressing a hybrid insulin gene-oncogene. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9037–9041. [PMC free article] [PubMed]
  • Mellon PL, Windle JJ, Goldsmith PC, Padula CA, Roberts JL, Weiner RI. Immortalization of hypothalamic GnRH neurons by genetically targeted tumorigenesis. Neuron. 1990 Jul;5(1):1–10. [PubMed]
  • Hammang JP, Baetge EE, Behringer RR, Brinster RL, Palmiter RD, Messing A. Immortalized retinal neurons derived from SV40 T-antigen-induced tumors in transgenic mice. Neuron. 1990 May;4(5):775–782. [PubMed]
  • Harlow E, Crawford LV, Pim DC, Williamson NM. Monoclonal antibodies specific for simian virus 40 tumor antigens. J Virol. 1981 Sep;39(3):861–869. [PMC free article] [PubMed]
  • Lane EB. Monoclonal antibodies provide specific intramolecular markers for the study of epithelial tonofilament organization. J Cell Biol. 1982 Mar;92(3):665–673. [PMC free article] [PubMed]
  • Botteri FM, van der Putten H, Wong DF, Sauvage CA, Evans RM. Unexpected thymic hyperplasia in transgenic mice harboring a neuronal promoter fused with simian virus 40 large T antigen. Mol Cell Biol. 1987 Sep;7(9):3178–3184. [PMC free article] [PubMed]
  • Brinster RL, Chen HY, Messing A, van Dyke T, Levine AJ, Palmiter RD. Transgenic mice harboring SV40 T-antigen genes develop characteristic brain tumors. Cell. 1984 Jun;37(2):367–379. [PubMed]
  • Spanopoulou E, Early A, Elliott J, Crispe N, Ladyman H, Ritter M, Watt S, Grosveld F, Kioussis D. Complex lymphoid and epithelial thymic tumours in Thy1-myc transgenic mice. Nature. 1989 Nov 9;342(6246):185–189. [PubMed]
  • Rüther U, Müller W, Sumida T, Tokuhisa T, Rajewsky K, Wagner EF. c-fos expression interferes with thymus development in transgenic mice. Cell. 1988 Jun 17;53(6):847–856. [PubMed]

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