4-Epidoxycycline: an alternative to doxycycline to control gene expression in conditional mouse models

Eger, Kurt; Hermes, Matthias; Uhlemann, Kathrin; Rodewald, Sabrina; Ortwein, J.; Brulport, Marc; Bauer, Alexander; Schormann, Wiebke; Lupatsch, F.; Schiffer, Ilka B.; Heimerdinger, Carolin K.; Gebhard, Susanne; Spangenberg, Christian; Prawitt, Dirk; Trost, Tatjana M.; Zabel, Bernhard; Sauer, Curtis M.; Berno, Tamara; Kölbl, H.; Krügel, Undine; Franke, Helmut; Illéš, Peter; Madaj-Sterba, P.; Bockamp, Ernesto; Beckers, Thomas; Hengstler, Jan G.

doi:10.1016/j.bbrc.2004.08.187

Cited by 22 publications

(23 citation statements)

References 24 publications

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“…In the future, the design of more efficient drugs may help enhance the efficacy of these expression systems. For instance, 4-epidoxycycline (4-ED), a hepatic metabolite of dox without antimicrobial activity or side effects on intestinal flora, was recently shown to be an efficient alternative to dox for regulating transgene expression in mice (Eger et al, 2004). A new induction system using photolabile-caged dox was also developed for cell-specific delivery and local gene expression that may be useful in vivo (Cambridge et al, 2004).…”

Section: Fig 1 Amentioning

confidence: 98%

Inducible and neuron‐specific gene expression in the adult mouse brain with the rtTA2S‐M2 system

Michalon

Koshibu

Baumgärtel

et al. 2005

Genesis

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To achieve inducible and reversible gene expression in the adult mouse brain, we exploited an improved version of the tetracycline-controlled transactivator-based system (rtTA2(S)-M2, rtTA2 hereafter) and combined it with the forebrain-specific CaMKIIalpha promoter. Several independent lines of transgenic mice carrying the CaMKIIalpha promoter-rtTA2 gene were generated and examined for anatomical profile, doxycycline (dox)-dependence, time course, and reversibility of gene expression using several lacZ reporter lines. In two independent rtTA2-expressing lines, dox-treatment in the diet induced lacZ reporter expression in neurons of several forebrain structures including cortex, striatum, hippocampus, amygdala, and olfactory bulb. Gene expression was dose-dependent and was fully reversible. Further, a similar pattern of expression was obtained in three independent reporter lines, indicating the consistency of gene expression. Transgene expression could also be activated in the developing brain (P0) by dox-treatment of gestating females. These new rtTA2-expressing mice allowing inducible and reversible gene expression in the adult or developing forebrain represent useful models for future genetic studies of brain functions.

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Section: Fig 1 Amentioning

confidence: 98%

Inducible and neuron‐specific gene expression in the adult mouse brain with the rtTA2S‐M2 system

Michalon

Koshibu

Baumgärtel

et al. 2005

Genesis

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“…Tracking and tracing of transplanted cells in mice is required in many Welds of research (Lewin et al 2000;Voura et al 2004;von Mach et al 2004;SchiVer et al 2003;Mohrmann et al 2005;Trost et al 2005;Eger et al 2004). Tracking of individual cells can be achieved by three strategies: (1) expression of marker proteins, such as enhanced green Xuorescent protein (EGFP) or -galactosidase, (2) Xuorescent dyes, exogenously loaded into the cells (Zhang et al 2002) and (3) inorganic nanocrystals or nanoparticles (Koeppel et al 2007;Jaiswal et al 2004;Larson Fig.…”

Section: Discussionmentioning

confidence: 97%

Tracking of human cells in mice

Schormann

Hammersen

Brulport

et al. 2008

Histochem Cell Biol

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Tracking and tracing of transplanted cells in mice is required in many fields of research. Examples are transplantation of stem cells into organs of mice to study their differentiation capacity and injection of tumor cells to examine metastatic behavior. In the present study we tested the lipid dye CM-DiI and red fluorescent nanoparticles Qdot655 for their applicability in tagging and tracing of human cells in mice. Labeling of different cell types, including MCF-7 human breast cancer cells, human cord blood derived cells, human NeoHep cells and human hepatopancreatic precursor cells, is technically easy and did not compromise further cell culture. After transplantation of CM-DiI or Qdot655 marked cells, red fluorescent structures could be detected already in unprocessed paraffin slices of the studied organs, namely liver, lung, pancreas, kidney, spleen and bone marrow. Next, we examined whether the red fluorescent structures represent the transplanted human cells. For this purpose, we established an in situ hybridization (ISH) technique that allows clear-cut differentiation between human and murine nuclei, based on simultaneous hybridization with human alu and mouse major satellite (mms) probes. We observed a high degree of coincidence between CM-DiI-marked cells and alu positive nuclei. However, also some mms positive cells contained CM-DiI, suggesting phagocytosis of the transplanted CM-DiI-marked cells. The degree of such CM-DiI-positive mouse cells depended on the cell type and route of administration. From a technical point of view it was important that CM-DiI-positive structures in paraffin slices remained fluorescent also after ISH. In contrast, Qdot655 positive structures faded during further staining procedures. In conclusion, marking of cells with CM-DiI or Qdot655 prior to transplantation facilitates recovery of human cells, since a high fraction of positive structures in the host's tissue originate from the transplanted cells. However, CM-DiI or Qdot655 positive staining of individual cells in transplanted tissues is not sufficient to prove their human origin. Additional procedures, such as ISH with alu-probes, are essential, when characterizing individual cells.

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“…The side effects of prolonged administration of Dox include renal toxicity, dose-dependent photosensitivity, tooth decoloration and decreased bone growth rate [36] . This problem of potentially raising antibiotic resistance to Dox may be overcome by the use of 4-epidoxycycline, a hepatic metabolite of Dox lacking antibiotic activity in mice [37] .…”

Section: Discussionmentioning

confidence: 99%

Developing a System for Regulating Expression of Human Hepatocyte Growth Factor Using Tetracycline in NRK52E Cells

Ren

Yu²,

Zhang³

et al. 2010

Urol Int

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Introduction: Hepatocyte growth factor (HGF) is a target of gene therapy for renal fibrosis. The aim of this study was to establish a human HGF gene expression system that is regulated by tetracycline (Tet) in normal rat kidney tubular epithelial cells (NRK52E cells). Materials and Methods: The plasmids pTet-on, pBI-L-HGF and pTK-Hyg were transfected sequentially into NRK52E cells using Lipofectamine 2000. The expression of HGF gene was measured, and the activity of expressed HGF was detected. Results: A clone of pBI-L-HGF/NRK52E cells showing strong reaction to doxycycline (Dox) was selected using a luciferase reporter assay system. The expression of both HGF mRNA and protein was significantly higher (both p < 0.01) in the Dox group than that in the control group. Furthermore, the bioactivity of expressed HGF was confirmed in the assay. Conclusions: A Tet-regulated human HGF gene expression system in NRK52E cells has been established. This cell line may prove useful for gene therapy against renal fibrosis.

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4-Epidoxycycline: an alternative to doxycycline to control gene expression in conditional mouse models

Cited by 22 publications

References 24 publications

Inducible and neuron‐specific gene expression in the adult mouse brain with the rtTA2S‐M2 system

Inducible and neuron‐specific gene expression in the adult mouse brain with the rtTA2S‐M2 system

Tracking of human cells in mice

Developing a System for Regulating Expression of Human Hepatocyte Growth Factor Using Tetracycline in NRK52E Cells

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