Efficient Transfection of Chicken Blastoderms in vivo by Lipofection and Electroporation Using Green Fluorescent Protein Gene as a Marker

Abstract

Chicken blastoderms of freshly laid and unincubated eggs at stage X were transfected in vivo by lipofection and electroporation using green fluorescent protein (GFP) gene as a marker. DNA was injected into the blastoderm with or without lipofection solution. Electroporation was carried out using a pair of electrodes in parallel on both sides of the blastoderm. The manipulated embryos were cultured in host eggshells and the GFP gene expression was detected through the aperture of the reconstituted eggs under a fluorescent microscope. When blastoderms were transfected by lipofection, expression of the GFP gene tended to be detected in the whole blastoderm. When blastoderms were transfected by electroporation, the GFP gene tended to express intensely in a limited part of the blastoderm. By combining both lipofection and electroporation, transfection efficiency was enhanced and the GFP gene expression in the whole blastoderm was observed more intensely. Embryonic and extra-embryonic tissue expression of the GFP gene was observed in 2-3 day incubated embryos both by lipofection and electroporation. GFP gene is very useful as a marker for detecting the gene expression because GFP gene expression can continuously be monitored in developing live embryos cultured in host eggshells.