EXOGENOUS GENE TRANSFECTION INTO QUAIL EMBRYO USING CATIONIC LIPID VESICLES

We have previously reported a simple procedure for gene transfection m ediated by cationic lipid vesicles for animal cells, in which a commer cially available synthetic cationic surfactant, dimethyldioctadecyl am monium bromide (DDAB), was used for making lipid vesicles. The transfe ction method was associated with low cytotoxicity and high transfectio n efficiency. In the present study, the method was applied for gene tr ansfection into quail embryos. The complex solution of lipid vesicles and pmiwZ plasmid, a beta-galactosidase expression vector under the co ntrol of beta-actin promoter as a reporter, was injected into quail em bryos at various developmental stages using a glass micropipette. Afte r incubation at 37.7 degrees C for 3-4 d, the embryos were fixed and s tained with X-gal solution. Under optimal conditions, about 80% of qua il embryos expressed beta-galactosidase in limited regions of tissues and organs with high viability. Moreover, 35% of the treated embryos ( 15/43) hatched following in vitro embryo culture, using the method dev eloped by us.