Introduction of exogenous DNA into gonads of chick embryos by lipofection and electroporation of stage X blastoderms in vivo.
1. In order to introduce exogenous DNA into gonads of chick embryos, stage X blastoderms of freshly laid and unincubated eggs were transfected by lipofection and electroporation in vivo. 2. The introduced DNA, green fluorescence protein (GFP) gene, was efficiently expressed in the blastoderms incubated for 24 h (78.8%, 78/99). 3. The GFP gene was present in most of the embryonic bodies and extra-embryonic membranes died by d 10 of incubation, when analysed by polymerase chain reaction. On d 16 to 20 of incubation, the GFP gene was detected in 7.0 to 20.9% of embryos in the heart, liver, stomach and brain, but not in the sartorius muscle. For the gonads, the GFP gene was detected in 22.2% (6/27) of the testes and 6.3% (1/18) of the ovaries examined. 4. These results suggest that it is possible to introduce exogenous DNA into gonads of chick embryos by lipofection and electroporation of stage X blastoderms in vivo.
Sano
A
A
Animal Genetic Engineering Laboratory, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan. mnaito@affrc.go.jp
Tagami
T
T
Harumi
T
T
Matsubara
Y
Y
Naito
M
M
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D001756:Blastoderm / Q000502:physiology*; D002642:Chick Embryo / Q000502:physiology*; D004247:DNA / Q000235:genetics*; D017931:DNA Primers; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics*; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000032:analysis; D014162:Transfection / Q000379:methods
D017931:DNA Primers; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D004247:DNA