Lentivirally generated eGFP-transgenic rats allow efficient cell tracking in vivo
Jens van den Brandt
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Search for more papers by this authorDapeng Wang
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Search for more papers by this authorSoon-Hwan Kwon
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Search for more papers by this authorMartin Heinkelein
Department of Virology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Search for more papers by this authorCorresponding Author
Holger M. Reichardt
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Versbacher Strasse 7, 97078 Würzburg, GermanySearch for more papers by this authorJens van den Brandt
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Search for more papers by this authorDapeng Wang
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Search for more papers by this authorSoon-Hwan Kwon
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Search for more papers by this authorMartin Heinkelein
Department of Virology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Search for more papers by this authorCorresponding Author
Holger M. Reichardt
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Versbacher Strasse 7, 97078 Würzburg, GermanySearch for more papers by this authorAbstract
Here we describe the efficient generation of eGFP-transgenic rats using a lentiviral approach. Analysis of the founder generation demonstrated that 46% of the offspring had stably integrated the provirus into the genome and of those 92% expressed eGFP in all blood-derived leukocytes. In contrast to their offspring, all founder rats were mosaic with regard to eGFP-expression, suggesting delayed viral transduction after injection. The expression level of eGFP in the F1 generation is influenced by and segregates with the site of proviral integration. Interestingly, a single copy of the transgene is sufficient for reliable detection by flow cytometry, irrespective of the leukocyte subtype analyzed. Adoptive transfer of purified CD4+ T-lymphocytes from transgenic rats and subsequent reisolation from various organs further demonstrated that expression of the lentiviral transgene is maintained in a foreign host and therefore allows for efficient tracking of transferred cells. Taken together, lentivirally generated eGFP-transgenic rats are a powerful tool for various applications in immunology and presumably also many other fields. genesis 39:94–99, 2004. © 2004 Wiley-Liss, Inc.
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