Original Paper

Transgenic Research

, Volume 19, Issue 3, pp 499-509

First online:

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

  • Qian ZhangAffiliated withEppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, 985950 Nebraska Medical Center
  • , Aleata A. TriplettAffiliated withEppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, 985950 Nebraska Medical Center
  • , Don W. HarmsAffiliated withMouse Genome Engineering Core Facility, Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center
  • , Wan-Chi LinAffiliated withEppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, 985950 Nebraska Medical Center
  • , Bradley A. CreamerAffiliated withEppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, 985950 Nebraska Medical Center
  • , Angie RizzinoAffiliated withEppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, 985950 Nebraska Medical CenterDepartment of Pathology and Microbiology, University of Nebraska Medical Center, 985950 Nebraska Medical Center
  • , Kay-Uwe WagnerAffiliated withEppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, 985950 Nebraska Medical CenterDepartment of Pathology and Microbiology, University of Nebraska Medical Center, 985950 Nebraska Medical Center Email author 

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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