Gene Transfer into Sheep

Gene transfer into animals has considerable potential for livestock improvement. If this potential is to be realized, the ease of generation of transgenic livestock will be of major importance. We report here the production of six transgenic sheep by microinjection of DNA into early embryos (1.2% of embryos transferred). Three different gene constructs were injected and transgenic sheep were obtained with each. The transgenic animals have all incorporated the DNA without detectable rearrangement, and where multiple copies were integrated, they are present in arrays of tandem repeats. Transmission of transferred genes to progeny of three of the sheep has been demonstrated. Five founder transgenic sheep described carry genes designed to direct the production of human clotting factor IX or human l−antitrypsin in milk. Transgenic animals carrying such genes may ultimately provide a new source of these and other therapeutic proteins.

1. Palmiter, R.D. and Brinster, R.L. 1986. Germline transformation of mice. Ann. Rev. Genet. 20:465−499. | Article | PubMed  | ISI | ChemPort |
2. Palmiter, R.D., Brinster, R.L., Hammer, R.E., Trumbauer, M.E., Rosenfeld, M.G., Birnberg, N.C., and Evans, R.M. 1982. Dramatic growth of mice that develop from eggs microinjected with metallothionein-growth hormone fusion genes. Nature 300:611−615. | PubMed  | ISI | ChemPort |
3. Palmiter, R.D., Norstedt, G., Gelinas, R.E., Hammer, R.E., and Brinster, R.L. 1983. Metallothionein-human GH fusion genes stimulate growth of mice. Science 222:809−814. | PubMed  | ISI | ChemPort |
4. Morello, D., Moore, G., Salmon, A.M., Yaniv, M., and Babinet, C. 1986. Studies on the expression of an H-2K/human growth hormone fusion gene in giant transgenic mice. EMBO J. 5:1877−1883. | PubMed  | ISI | ChemPort |
5. Hammer, R.E., Pursel, V.G., Rexroad, C.E., Jr., Wall, R.J., Bolt, D.J., Ebert, K.M., Palmiter, R.D., and Brinster, R.L. 1985. Production of transgenic rabbits, sheep and pigs by microinjection. Nature 315:680−683. | PubMed  | ISI | ChemPort |
6. Evans, J.W. and Hollaender, A. 1986. Genetic Engineering of Animals. Plenum, New York.
7. Ward, K.A., Franklin, I.R., Murray, J.D., Nancarrow, C.D., Raphael, K.A., Rigby, N.W., Byrne, C.R., Wilson, B.W., and Hunt, C.L. 1986. The direct transfer of DNA by embryo microinjection. In: Proceedings 3rd World Congress Applied to Livestock Production, p. 6−21. G. E. Dickerson & R.K. Johnson, (eds.). University of Nebraska, Lincoln, Nebraska.
8. Simons, J.P. and Land, R.B. 1987. Transgenic Livestock, J. Reprod. Fertil. Suppl. 34:237−250. | PubMed  | ChemPort |
9. Clark, A.J., Simons, P., Wilmut, I., and Lathe, R. 1987. Pharmaceuticals from transgenic livestock. Trends Biotechnol. 5:20−24. | ISI | ChemPort |
10. Lathe, R., Clark, A.J., Archibald, A.L., Bishop, J.O., Simons, P., and Wilmut, I. 1986. Novel products from livestock. In: Exploiting new technologies in animal breeding: genetic developments. Smith, C., King, J.W.B., and McKay, J.C. (eds.). Oxford University Press, Oxford.
11. Simons, J.P., McClenaghan, M., and Clark, A.J. 1987. Alteration of the quality of milk by expression of sheep -lactoglobulin in transgenic mice. Nature 328:530−532. | Article | PubMed  | ISI | ChemPort |
12. Gordon, K., Lee, E., Vitale, J.A., Smith, A.E., Westphal, H., and Hennighausen, L. 1987. Production of human tissue plasminogen activator in transgenic mouse milk. Bio Technology 5:1183−1187. | ISI | ChemPort |
13. Brem, G., Brenig, B., Goodman, H.M., Selden, R.C., Graf, F., Kruff, B., Springman, K., Hondele, J., Meyer, J., Winnacker, E.-L., and Kräulich, H. 1985. Zuchthygiene 20:251−252. | ISI |
14. Brinster, R.L., Chen, H.Y., Trumbauer, M.E., Yagle, M.K., and Palmiter, R.D. 1985. Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc. Natl. Acad. Sci. U.S.A. 82:4438−4442. | PubMed  | ChemPort |
15. Brinster, R.L., Chen, H.Y., and Trumbauer, M. 1981. Somatic expression of herpes thymidine kinase in mice following injection of a fusion gene into eggs. Cell 27:223−231. | Article | PubMed  | ISI | ChemPort |
16. Palmiter, R.D., Chen, H.Y., and Brinster, R.L. 1982. Differential regulation of metallothionein-thymidine kinase fusion genes in transgenic mice and their offspring. Cell 29:701−710. | Article | PubMed  | ISI | ChemPort |
17. Ali, S. and Clark, A.J. 1988. Characterisation of the gene encoding -lactoglobulin: similarity to the genes for retinol binding protein and other secretory proteins. J. Molec. Biol. In press. | PubMed  |
18. Chada, K., Magram, J., Raphael, K., Radice, G., Lacy, E., and Costantini, F. 1985. Specific expression of a foreign -globin gene in erythroid cells of transgenic mice. Nature 314:377−380. | PubMed  | ISI | ChemPort |
19. Townes, T.M., Lingrel, J.B., Chen, H.Y., Brinster, R.L., and Palmiter, R.D. 1985. Erythroid-specific expression of human -globin genes in transgenic mice. EMBO J. 4:1715−1723. | PubMed  | ISI | ChemPort |
20. Hammer, R.E., Krumlauf, R., Camper, S.A., Brinster, R.L., and Tilghman, S.M. 1987. Diversity of alpha-fetoprotein gene expression in mice is generated by a combination of separate enhancer elements. Science 235:53−58. | PubMed  | ISI | ChemPort |
21. Pursel, V.G., Miller, K.F., Pinkert, C.A., Palmiter, R.D., and Brinster, R.L. 1987. Development of 1-cell and 2-cell pig ova after microinjection of genes. J. Anim. Sci. 65 (Suppl.1):402 (Abstr.).
22. Hammer, R.E., Palmiter, R.D., and Brinster, R.L. 1984. Partial correction of murine hereditary growth disorder by germ-line incorporation of a new gene. Nature 311:65−67. | PubMed  | ISI | ChemPort |
23. Wilkie, T.M., Brinster, R.L., and Palmiter, R.D. 1986. Germline and somatic mosaicism in transgenic mice. Devel. Biol. 118:9−18. | ISI | ChemPort |
24. Stockwell Hartree, A., Mills, J.B., Welch, R., and Thomas, M. 1968. Fractionation of protein hormones from horse pituitary glands. J. Reprod. Fertil. 17:291−303. | PubMed  |
25. Hunter, G.L., Adams, C.E., and Rowson, L.E.A. 1955. Interbreed ovum transfer in sheep. J. Agric. Sci. 46:143−149.
26. Hogan, B., Costantini, F., and Lacy, E. 1986. Manipulating the Mouse Embryo: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
27. Anson, D., Austen, D.E.G., and Brownlee, G.G. 1985. Expression of active human clotting factor IX from recombinant DNA clones in mammalian cells. Nature 315:683−685. | PubMed  | ISI | ChemPort |
28. Lathe, R., Vilotte, J.-L., and Clark, A.J. 1987. Plasmid and bacteriophage vectors for excision of intact inserts. Gene 57:193−201. | Article | PubMed  | ISI | ChemPort |
29. Spandidos, D.A., and Paul, J. 1982. Transfer of human globin genes to erythroleukemic mouse cells. EMBO J. 1:15−20. | PubMed  | ISI | ChemPort |
30. Gaye, P., Hue-Delahaie, D., Mercier, J.-C., Soulier, S., Vilotte, J.-L., and Furet, J.-P. 1986. Ovine -lactoglobulin messenger RNA: nucleo-tide sequence and mRNA levels during functional differentiation of the mammary gland. Biochimie 68:1097−1107. | PubMed  | ISI | ChemPort |

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