Transgenic Research

, Volume 19, Issue 3, pp 499–509

Temporally and spatially controlled expression of transgenes in embryonic and adult tissues

  • Qian Zhang
  • Aleata A. Triplett
  • Don W. Harms
  • Wan-Chi Lin
  • Bradley A. Creamer
  • Angie Rizzino
  • Kay-Uwe Wagner
Original Paper

Abstract

Using ES cell-mediated transgenesis, we generated a novel mouse strain that permits a temporally and spatially controlled expression of responder genes in embryonic and multiple adult tissues. The transgene was constructed in a way that a CMV enhancer linked to the chicken β-actin promoter (CAG) drives the expression of the tetracycline-controlled transactivator (tTA) in particular tissues upon Cre-mediated excision of a floxed βgeo marker located between the promoter and the tTA. Based on the enzymatic activity of lacZ, the CAG-βgeo-tTA construct exhibits a widespread expression and appears to be very strong in the brain, heart, muscle, pancreas, and skin. Like the embryonic stem cell line that was used to generate this strain, the CAG-βgeo-tTA transgene is already highly active in preimplantation embryos. Using in vivo bioluminescence imaging on MMTV-Cre, CAG-βgeo-tTA, TetO-Luciferase triple transgenic mice and their controls, we demonstrated that the expression of the tTA, which is strictly dependent on the presence of Cre recombinase, induces the activation of the reporter transgene in the absence of any ligands. The tTA-mediated transactivation can be completely ablated through administration of doxycycline, and its subsequent withdrawal lifts the transcriptional block. Based on these characteristics, this novel strain may be useful in experiments that require a sustained expression of transgenes in particular cell types over a prolonged period followed by a rapid downregulation, for example in studies that examine the therapeutic value of cancer-initiating oncogenes during disease progression.

Keywords

Gene expression regulation Transgenic Stem cells Cre recombinase Tetracycline Trans-activators 

Supplementary material

11248_2009_9329_MOESM1_ESM.tif (7.2 mb)
Supplementary material (TIFF 7403 kb)

References

  1. Adams JM, Harris AW, Pinkert CA, Corcoran LM, Alexander WS, Cory S, Palmiter RD, Brinster RL (1985) The c-myc oncogene driven by immunoglobulin enhancers induces lymphoid malignancy in transgenic mice. Nature 318:533–538CrossRefPubMedGoogle Scholar
  2. Belteki G, Haigh J, Kabacs N, Haigh K, Sison K, Costantini F, Whitsett J, Quaggin SE, Nagy A (2005) Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction. Nucleic Acids Res 33:e51CrossRefPubMedGoogle Scholar
  3. Brinster RL, Chen HY, Messing A, Van Dyke T, Levine AJ, Palmiter RD (1984) Transgenic mice harboring SV40 T-antigen genes develop characteristic brain tumors. Cell 37:367–379CrossRefPubMedGoogle Scholar
  4. Creamer BA, Triplett AA, Wagner KU (2009) Longitudinal analysis of mammogenesis using a novel tetracycline-inducible mouse model and in vivo imaging. Genesis 47:234–245CrossRefPubMedGoogle Scholar
  5. Ewald D, Li M, Efrat S, Auer G, Wall RJ, Furth PA, Hennighausen L (1996) Time-sensitive reversal of hyperplasia in transgenic mice expressing SV40 T antigen. Science 273:1384–1386CrossRefPubMedGoogle Scholar
  6. Furth PA, St Onge L, Boger H, Gruss P, Gossen M, Kistner A, Bujard H, Hennighausen L (1994) Temporal control of gene expression in transgenic mice by a tetracycline-responsive promoter. Proc Natl Acad Sci USA 91:9302–9306CrossRefPubMedGoogle Scholar
  7. Gossen M, Bujard H (1992) Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci USA 89:5547–5551CrossRefPubMedGoogle Scholar
  8. Hennighausen L, Wall RJ, Tillmann U, Li M, Furth PA (1995) Conditional gene expression in secretory tissues and skin of transgenic mice using the MMTV-LTR and the tetracycline responsive system. J Cell Biochem 59:463–472CrossRefPubMedGoogle Scholar
  9. Kelly DL, Rizzino A (2000) DNA microarray analyses of genes regulated during the differentiation of embryonic stem cells. Mol Reprod Dev 56:113–123CrossRefPubMedGoogle Scholar
  10. Lobe CG, Koop KE, Kreppner W, Lomeli H, Gertsenstein M, Nagy A (1999) Z/AP, a double reporter for cre-mediated recombination. Dev Biol 208:281–292CrossRefPubMedGoogle Scholar
  11. Mayford M, Bach ME, Huang YY, Wang L, Hawkins RD, Kandel ER (1996) Control of memory formation through regulated expression of a CaMKII transgene. Science 274:1678–1683CrossRefPubMedGoogle Scholar
  12. Stewart TA, Pattengale PK, Leder P (1984) Spontaneous mammary adenocarcinomas in transgenic mice that carry and express MTV/myc fusion genes. Cell 38:627–637CrossRefPubMedGoogle Scholar
  13. Wagner KU, Wall RJ, St-Onge L, Gruss P, Wynshaw-Boris A, Garrett L, Li M, Furth PA, Hennighausen L (1997) Cre-mediated gene deletion in the mammary gland. Nucleic Acids Res 25:4323–4330CrossRefPubMedGoogle Scholar
  14. Wagner KU, Claudio E, Rucker EB III, Riedlinger G, Broussard C, Schwartzberg PL, Siebenlist U, Hennighausen L (2000) Conditional deletion of the bcl-x gene from erythroid cells results in hemolytic anemia and profound splenomegaly. Development 127:4949–4958PubMedGoogle Scholar
  15. Wagner KU, McAllister K, Ward T, Davis B, Wiseman R, Hennighausen L (2001) Spatial and temporal expression of the Cre gene under the control of the MMTV-LTR in different lines of transgenic mice. Transgenic Res 10:545–553CrossRefPubMedGoogle Scholar
  16. Wagner KU, Krempler A, Triplett AA, Qi Y, George NM, Zhu J, Rui H (2004) Impaired alveologenesis and maintenance of secretory mammary epithelial cells in Jak2 conditional knockout mice. Mol Cell Biol 24:5510–5520CrossRefPubMedGoogle Scholar
  17. Yu HM, Liu B, Chiu SY, Costantini F, Hsu W (2005) Development of a unique system for spatiotemporal and lineage-specific gene expression in mice. Proc Natl Acad Sci USA 102:8615–8620CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Qian Zhang
    • 1
  • Aleata A. Triplett
    • 1
  • Don W. Harms
    • 2
  • Wan-Chi Lin
    • 1
  • Bradley A. Creamer
    • 1
  • Angie Rizzino
    • 1
    • 3
  • Kay-Uwe Wagner
    • 1
    • 3
  1. 1.Eppley Institute for Research in Cancer and Allied DiseasesUniversity of Nebraska Medical Center, 985950 Nebraska Medical CenterOmahaUSA
  2. 2.Mouse Genome Engineering Core Facility, Department of Genetics, Cell Biology and AnatomyUniversity of Nebraska Medical CenterOmahaUSA
  3. 3.Department of Pathology and MicrobiologyUniversity of Nebraska Medical Center, 985950 Nebraska Medical CenterOmahaUSA

Personalised recommendations