Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Production of goats by somatic cell nuclear transfer

Nature Biotechnology volume 17, pages 456–461 (1999)Cite this article

Abstract

In this study, we demonstrate the production of transgenic goats by nuclear transfer of fetal somatic cells. Donor karyoplasts were obtained from a primary fetal somatic cell line derived from a 40-day transgenic female fetus produced by artificial insemination of a nontransgenic adult female with semen from a transgenic male. Live offspring were produced with two nuclear transfer procedures. In one protocol, oocytes at the arrested metaphase II stage were enucleated, electrofused with donor somatic cells, and simultaneously activated. In the second protocol, activated in vivo oocytes were enucleated at the telophase II stage, electrofused with donor somatic cells, and simultaneously activated a second time to induce genome reactivation. Three healthy identical female offspring were born. Genotypic analyses confirmed that all cloned offspring were derived from the donor cell line. Analysis of the milk of one of the transgenic cloned animals showed high-level production of human antithrombin III, similar to the parental transgenic line.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: (A) In vitro-cultured caprine fetal somatic cells at Passage 6, magnification ×200.
Figure 2: Genotypic analysis of nuclear transfer goats.
Figure 3: Western blot of rhAT expressed in the milk of the cloned transgenic doe CFF6-1.

References

  1. Meade, H.M. et al. in Gene expression systems: using nature for the art of expression, (eds Fernandez, J.M. & Hoeffler, J.P.) 399–427 (Academic Press, San Diego; 1998).

  2. Clark, A.J. The mammary gland as a bioreactor: expression, processing, and production of recombinant proteins. J. Mammary Gland Biol. Neoplasia 3, 337–350 (1998).

    Article  CAS  Google Scholar 

  3. Edmunds, T. et al. Transgenically produced human antithrombin—structural and functional comparison to human plasma-derived antithrombin. Blood 91, 4561–4571 (1998).

    CAS  PubMed  Google Scholar 

  4. Wineland, N.E., Detwiler, L.A. & Salman, M.D. Epidemiologic analysis of reported scrapie in sheep in the United States: 1,117 cases (1947–1992). J. Am. Vet. Med. Assoc. 212, 713–718 (1998).

  5. Ebert, K.M. & Schindler, J.E.S. Transgenic farm animals: Progress report. Theriogenology 39, 121– 135 (1993).

    Article  Google Scholar 

  6. Wilkie, T.M., Brinster, R.L. & Palmiter, R.D. Germline and somatic mosaicism in transgenic mice. Dev. Biol. 118, 9–18 (1986).

    Article  CAS  Google Scholar 

  7. Burdon, T.G. & Wall, R.J. Fate of microinjected genes in preimplantation mouse embryos. Mol. Reprod. Dev. 33, 436 –442 (1992).

    Article  CAS  Google Scholar 

  8. Whitelaw, C.B.A., Springbett, A.J., Webster, J. & Clark, A.J. The majority of G0 transgenic mice are derived from mosaic embryos. Transgenic Res. 2, 29– 32 (1992).

    Article  Google Scholar 

  9. Yong, Z. & Yuqiang, L. Nuclear-cytoplasmic interaction and development of goat embryos reconstructed by nuclear transplantation: production of goats by serially cloning embryos. Biol. Reprod. 58, 266–269 (1998).

    Article  CAS  Google Scholar 

  10. Campbell, K.H.S., McWhir, J., Ritchie, W.A. & Wilmut, I. Sheep cloned by nuclear transfer from a cultured cell line. Nature 380, 64–66, (1996).

    Article  CAS  Google Scholar 

  11. Wilmut, I., Schnieke, A.E., McWhir, J., Kind, A.J. & Campbell, K.H.S. Viable offspring derived from fetal and adult mammalian cells. Nature 385, 810–813 (1997).

    Article  CAS  Google Scholar 

  12. Wells, D.N., Misica, P.M., Day, T.A. & Tervit, H.R. Production of cloned lambs from an established embryonic cell line: a comparison between in vivo- and in vitro-matured cytoplasts. Biol. Reprod 57, 385–393 (1997).

    Article  CAS  Google Scholar 

  13. Schnieke, A.E. et al. Human Factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts. Science 278, 2130–2133 (1997).

    Article  CAS  Google Scholar 

  14. Cibelli, J.B. et al. Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science 280, 1256–1258 (1998).

    Article  CAS  Google Scholar 

  15. Vignon, X. et al. Developmental potential of bovine embryos reconstructed from enucleated matured oocytes fused with cultured somatic cells. C.R. Acad. Sci. 321, 735–745 (1998).

    Article  CAS  Google Scholar 

  16. Wakayama, T., Perry, A.C.F., Zuccotti, M., Johnson, K.R., & Yanagimachi, R. Full-term development of mice from enucleated oocytes injected with cumulus cells nuclei. Nature 394, 369–374 (1998).

    Article  CAS  Google Scholar 

  17. Kato, Y. et al. Eight calves cloned form somatic cells of a single adult. Science 282, 2095–2098 (1998).

    Article  CAS  Google Scholar 

  18. Amills, M., Francino, O. & Sànchez, A. Nested PCR allows the characterization of TaqI and PstI RFLPs in the second exon of the caprine MHC class II DRB gene. Vet. Immunol. Immunopathol. 48, 313–321 (1996).

    Article  Google Scholar 

  19. Bordignon, V. & Smith, L.C. Telophase enucleation: an improved method to prepare recipient cytoplasts for use in bovine nuclear transfer. Mol. Reprod. Dev. 49, 29– 36 (1998).

    Article  CAS  Google Scholar 

  20. Laird, P.W. et al. Simplified mammalian DNA isolation procedure. Nucleic Acids Res. 19, 4293 (1991).

    Article  CAS  Google Scholar 

  21. Aasen, E. & Medrano, J.F. Amplification of the ZFY and ZFX genes for sex identification in humans, cattle, sheep and goats. Bio/Technology 8, 1279– 1281 (1990).

    CAS  PubMed  Google Scholar 

  22. Gavin, W.G. in Transgenic animals—generation and use (ed. Houdebine, L.M.) 19–21 (Harwood Academic Publishers, Amsterdam, 1996).

    Google Scholar 

  23. Maniatis, T., Fritsch, E.F. & Sambrook, J. Molecular cloning, a laboratory manual. (Cold Spring Harbor Press, Cold Spring Harbor, NY, 1982).

    Google Scholar 

  24. Church, G.M. & Gilbert, W. Genomic sequencing. Proc. Natl. Acad. Sci. USA 81, 1991– 1995 (1984).

    Article  CAS  Google Scholar 

  25. Amills, M., Francino, O. & Sànchez, A. A PCR-RFLP typing method for the caprine Mhc class II DRB gene. Vet. Immunol. Immunopathol. 55, 255–260 (1996).

    Article  CAS  Google Scholar 

  26. Ponce de Leon, F.A., Li, Y. & Weng, Z. Early and late replicative chromosomal banding patterns of Gallus domesticus. J. Hered. 83, 36– 42 (1992).

    Article  CAS  Google Scholar 

  27. van de Corput, M.P.C., Dirks, R.W., van Gijlswijk, R.P.M., van de Rijke, F.M. & Raap, A.K. Fluorescence in situ hybridization using horseradish peroxidase-labeled oligodeoxynucleotides and tyramide signal amplification for sensitive DNA and mRNA detection. Histochem Cell Biol. 110, 431– 437 (1998).

    Article  CAS  Google Scholar 

  28. Klinger, K. et al. Rapid detection of chromosome aneuploidies in uncultured amniocytes by using fluorescence in situ hybridization (FISH). Am. J. Hum. Genet. 51, 55–65 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Di Berardino, D. et al. R-banding pattern of the prometaphase chromosomes of the goat. J. Hered. 78, 225–230 (1987).

    Article  CAS  Google Scholar 

  30. Ryot, K.D., Vadnere S.V. & Prakash P. Hormonal induction of lactation and histomorphology of mammary glands in prepubertal goats. Indian J. Anim. Res. 10, 49–51 (1989).

    Google Scholar 

Download references

Acknowledgements

This work was supported, in part, by a grant from the NIH (Small Business Innovation Research Program, R43 HD35395-01) to Genzyme Transgenics Corporation, and by Genzyme Transgenics Corporation. The authors gratefully acknowledge the help of M. Schofield, D.V.M., A. O'Coin, S. Bombard, N. Hawkins, S. Blash, R. Burns, and B. Kuehholzer. We wish to thank Gary Anderson for critical reading of the manuscript and helpful comments.

Author information

Author notes

  1. Alexander Baguisi and Esmail Behboodi: These authors contributed equally to this work.

Authors and Affiliations

  1. Genzyme Transgenics Corporation, Framingham , 01701-9322, MA

    Esmail Behboodi, David T. Melican, Margaret M. Destrempes, Christine Cammuso, Jennifer L. Williams, Scott D. Nims, Catherine A. Porter, Patricia Midura, Carol A. Ziomek, Harry M. Meade, William G. Gavin & Yann Echelard

  2. Nuclear Transfer Laboratory, North Grafton , 01356, MA

    Alexander Baguisi, Monica J. Palacios & Eric W. Overström

  3. Department of Biomedical Sciences, Tufts University School of Veterinary Medicine, North Grafton, 01356, MA

    Alexander Baguisi, Monica J. Palacios, Sandra L. Ayres & Eric W. Overström

  4. Cell and Protein Therapeutic Department, Genzyme Corporation, Framingham, 01701-9322, MA

    Julie S. Pollock & Michael L. Hayes

  5. Department of Animal Science, Lousiana State University, Baton Rouge, 70803, LA

    Richard S. Denniston & Robert A. Godke

Authors
  1. Alexander Baguisi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Esmail Behboodi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. David T. Melican
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Julie S. Pollock
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Margaret M. Destrempes
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Christine Cammuso
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jennifer L. Williams
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Scott D. Nims
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Catherine A. Porter
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Patricia Midura
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Monica J. Palacios
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Sandra L. Ayres
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Richard S. Denniston
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Michael L. Hayes
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Carol A. Ziomek
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Harry M. Meade
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Robert A. Godke
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. William G. Gavin
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Eric W. Overström
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. Yann Echelard
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Eric W. Overström or Yann Echelard.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Baguisi, A., Behboodi, E., Melican, D. et al. Production of goats by somatic cell nuclear transfer. Nat Biotechnol 17, 456–461 (1999). https://doi.org/10.1038/8632

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/8632

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing