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Application of green fluorescent protein as a marker for selection of transgenic mouse embryos before implantation

In this study, we evaluated the application of green fluorescent protein (GFP) from the jellyfish Aequorea victoria, for efficient selection of possible transgenic and non-mosaic mouse embryos after microinjection. | Turk J Biol 25 (2001) 123-131 © TÜB‹TAK Application of Green Fluorescent Protein as a Marker for Selection of Transgenic Mouse Embryos before Implantation Haydar BA⁄Ifi TÜB‹TAK, Marmara Research Center, Research Institute for Genetic Engineering and Biotechnology, P.O. Box 41, 41470 Gebze, Kocaeli-TURKEY Levent KESK‹NTEPE Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta GA 30912-2600, USA. Received: 17.09.1999 Abstract: In this study, we evaluated the application of green fluorescent protein (GFP) from the jellyfish Aequorea victoria, for efficient selection of possible transgenic and non-mosaic mouse embryos after microinjection. We injected 353 one-cell mouse zygotes with a DNA fragment carrying Gfp gene under the control of ß-actin gene promoter. Eighty-seven per cent (307 embryos) of the injected embryos survived after microinjection. The surviving embryos were cultured for an additional day and then analysed for detection of GFP expression using a standard fluorescein isothiocyanate (FITC) filter set attached to a microscope. In this analysis, 34 % of the two-cell stage embryos (37 embryos out of 108) were found to be GFP positive. Of these GFP-positive embryos, 8 embryos out of 37 (21%) were found to be non-mosaic as GFP expression was detected in all the blastomeres. Our results show that the successful use of GFP as a vital marker for detection of mosaic embryos holds promise for potential application of this technology as a selection tool in the generation of transgenic animals. Key Words: Transgenic mice, embryo, microinjection, green fluorescent protein Transgenik Fare Embriyolar›n›n ‹mplantasyon Öncesi Seçiminde Yeflil Floresans Proteini'nin Mark›r Olarak Kullan›m› Özet: Bu çal›flmada, Aequrea victoria (deniz anasi) dan elde edilen yeflil floresan proteinin (GFP) ekspresyonunun mark›r olarak kullan›lma olana¤›, mikroenjeksiyon sonras› elde edilen transgenik ve mozaik olmayan fare embriyolar›nda araflt›r›ld›. Bu .