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Title: PRIMORDIAL GERM CELLS FOR GENETIC MODIFICATION OF POULTRY

Author
item WENTWORTH, B - UNIV OF WISCONSIN
item TSAI, H - UNIV OF WISCONSIN
item WENTWORTH, A - UNIV OF WISCONSIN
item WONG, E - VIRGINIA POLYTECHNIC INST
item Proudman, John
item EL HALAWANI, M - UNIV OF MINNESOTA

Submitted to: Beltsville Agricultural Research Center Symposium
Publication Type: Review Article
Publication Acceptance Date: 2/22/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary: The production of transgenic animals offers substantial utility in agriculture for the improvement of productive traits and the enhancement of nutrient properties. Transgenic poultry will have unparalled product usefulness and will also provide enormous research potential for studying avian biological development, as well as endocrinological, cellular and molecular mechanisms of avian growth, reproduction and immunity. However, the technology to produce transgenic poultry lags behind that of transgenic mammals. Distinct features of the avian reproductive system necessitate original techniques to introduce exogenous DNA into the genome of the bird. This review describes the isolation, culture, and transfection of avian primordial germ cells (PGCs); transfer of PGCs transfected with foreign DNA into the blood vascular system of recipient embryos, and recovery of the transgene in the injected founder animals as well as their subsequent transgenic progeny. Also discussed are techniques for enhancement of transgenic efficiency by production of sterile recipients and the establishment of immortal blastodermal cell lines. Other methods of gene transfer in the avian species, and the implications of transgenic research, are also discussed.

Technical Abstract: Research has been directed toward the development in culture of five independent chicken blastodermal cell strains each of a single sex; transfection of cultured turkey gonadal primordial germ cells (gPGCs) with foreign DNA (lacZ or antisense prolactin DNA); transfer of these transfected gPGCs into the blood vascular system of stage 16-17 recipient embryos, and recovery of the DNA transgenes from the gonads of founder (F0 animals and from the muscle and blood of the F1 and F2 progeny of the mated recipients. The transfection efficiency of primary cultured gPGCs with either lacZ or antisense prolactin (asPRL) DNA using Lipofectamine as the carrier was 26%. The exogenous DNA was detected in the stage 36 F0 embryonic gonad by slot blot for lacZ using a P-32 probe for lacZ or by PCR and/or Southern blot using primers for a combined ASVLTR-asPRL fragment or a nick translated P-32 probe for asPRL, respectively. This non-viral method dof gene transfer in the avian species appears worthy of further investigation. Methods of transgenic enhancement by the production of sterile recipients (lacking germ cells) are also presented. The injection of embryonated chicken eggs with EMA-1 antibody and the immunization of hens with germ cell-like 500 Kd antigen resulted in sterile host embryos. Germ cells from the germinal crescent of quail embryos were able to populate the gonads of antibody sterilized host embryos. Other methods of gene transfer in the avian species and the implications of transgenic research are discussed.