Nature Biotechnology
17, 456 - 461 (1999)
doi:10.1038/8632
Production of goats by somatic cell nuclear transferAlexander Baguisi2, 3, 6, Esmail Behboodi1, 6, David T. Melican1, Julie S. Pollock4, Margaret M. Destrempes1, Christine Cammuso1, Jennifer L. Williams1, Scott D. Nims1, Catherine A. Porter1, Patricia Midura1, Monica J. Palacios2, 3, Sandra L. Ayres3, Richard S. Denniston5, Michael L. Hayes4, Carol A. Ziomek1, Harry M. Meade1, Robert A. Godke5, William G. Gavin1, Eric W. Overström2, 3
& Yann Echelard11
Genzyme Transgenics Corporation, Framingham
MA, 01701-9322. 2
Nuclear Transfer Laboratory, North Grafton
, MA 01356 and 3
Department of Biomedical Sciences, Tufts University
School of Veterinary Medicine, North Grafton, MA
01356. 4
Cell and Protein Therapeutic Department, Genzyme Corporation
, Framingham, MA 01701-9322. 5
Department of Animal Science, Lousiana State University
, Baton Rouge, LA 70803. 6
These authors contributed equally to this work.
Correspondence should be addressed to Eric W. Overström eoverstrom@infonet.tufts.edu or Yann Echelard yechelard@genzyme.com.nuclear transferoocytetransgenicantithrombin IIIgoatIn 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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