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Generation and characterization of a GFP transgenic rat line for embryological research

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Abstract

Model organisms expressing fluorescent proteins are important tools for research. The present study was performed to generate and characterize a new line of green fluorescent protein (GFP) transgenic rats for use as a model in experimental embryological research. We injected a GFP expression vector into 135 zygotes of the Sprague-Dawley (SD) rat strain. Embryo transfer of 103 surviving embryos resulted in the production of 35 offspring (33.9%) and two of them were transgenic (5.7%). Two transgenic rat lines that ubiquitously express GFP under the control of the cytomegalovirus-enhancer/β-actin (CAGGS) promoter were generated by breeding. We studied the main embryological parameters of one these GFP transgenic lines. Homozygous GFP-transgenic females have the same ovulation and superovulation rates as wild type (WT) females. Transgenic embryos reached blastocyst stage in vitro and developed in vivo after embryo transfer without decrease in their developmental ability compared to the control group. The genotype of the parents determined the onset of GFP expression in preimplantation embryos. When the GFP gene is derived from the transgenic female parent, fluorescence was detected in oocytes and in embryos of all further stages of development. When the GFP gene is inherited by the transgenic male parent, GFP was only expressed from the blastocyst stage on. GFP-transgenic rats represent a valuable tool to mark embryos for many embryological studies such as transgenesis, gene expression patterns during early development, embryo aggregation for analysis of the distribution of cells in chimeric embryos and nuclear transfer to confirm the origin of the cloned offspring.

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Acknowledgments

We like to thank Rosemarie Barnow and Cathrin Rudolph for skillful technical assistance and Natalia Alenina for her professional help with the preparation of the figures. This work was supported by the EURATools integrated project funded by the Sixth Framework Programme of the European Union.

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Authors and Affiliations

  1. Max-Delbrück Center for Molecular Medicine (MDC), Robert-Rössle-Str. 10, 13092, Berlin-Buch, Germany

    Elena Popova, Brit Rentzsch, Michael Bader & Alexander Krivokharchenko

  2. Department of Biology, Chemistry and Pharmacy, Free University Berlin, Takustr. 3, 14195, Berlin, Germany

    Brit Rentzsch

Authors
  1. Elena Popova
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  2. Brit Rentzsch
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  3. Michael Bader
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  4. Alexander Krivokharchenko
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Correspondence to Michael Bader.

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Popova, E., Rentzsch, B., Bader, M. et al. Generation and characterization of a GFP transgenic rat line for embryological research. Transgenic Res 17, 955–963 (2008). https://doi.org/10.1007/s11248-008-9189-0

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  • DOI: https://doi.org/10.1007/s11248-008-9189-0

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