Production of goats by somatic cell nuclear transfer

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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.

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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.

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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.

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Correspondence to Eric W. Overström or Yann Echelard.

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