Generation of transgenic mice by transfection of pronuclear embryos using lipid-DNA complexes.
We had previously developed a methodology to introduce foreign DNA into mouse eggs and embryos using cationic lipids as vectors. In this report we use this technique to produce transgenic animals. Mouse embryos at the pronuclear stage were transfected using a mixture of a plasmid DNA, encoding for a nuclear form of beta-galactosidase, and a commercial lipid transfection reagent. Embryos were washed and incubated overnight. Those that cleaved and develop to the 2-cell stage of normal appearance were transferred to the Fallopian tubes of pseudopregnant foster mothers. We analysed a total of 158 offspring and found two, a female and a male, to be transgenic (1.27% of the total). Integration of the foreign DNA in the female was showed by Southern blot. Both animals expressed the lacz in several organs, but none of them either displayed expression in the germ cells or transmitted the transgene to their offspring. Taken together our results show that lipid transfection can generate transgenic mice, but the efficiency needs to be improved for this method to be widely applied.
Carballada
Rosa
R
Centro de Investigaciones Biológicas, CSIC, Madrid, Spain.
Relloso
Miguel
M
Esponda
Pedro
P
D000818:Animals; D001483:Base Sequence; D004274:DNA, Recombinant / Q000008:administration & dosage* / Q000235:genetics*; D004622:Embryo, Mammalian; D005260:Female; D015870:Gene Expression; D017930:Genes, Reporter; D007763:Lac Operon; D008055:Lipids / Q000008:administration & dosage; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D014018:Tissue Distribution; D014162:Transfection / Q000379:methods*
D004274:DNA, Recombinant; D008055:Lipids