Production of goats by somatic cell nuclear transfer.
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.
Baguisi
A
A
Department of Biomedical Sciences, Tufts University School of Veterinary Medicine, North Grafton, MA 01356, USA.
Behboodi
E
E
Melican
D T
DT
Pollock
J S
JS
Destrempes
M M
MM
Cammuso
C
C
Williams
J L
JL
Nims
S D
SD
Porter
C A
CA
Midura
P
P
Palacios
M J
MJ
Ayres
S L
SL
Denniston
R S
RS
Hayes
M L
ML
Ziomek
C A
CA
Meade
H M
HM
Godke
R A
RA
Gavin
W G
WG
Overström
E W
EW
Echelard
Y
Y
D000818:Animals; D030801:Animals, Genetically Modified; D000990:Antithrombin III / Q000235:genetics; D015139:Blotting, Southern; D002467:Cell Nucleus / Q000502:physiology; D019976:Cloning, Organism; D005314:Embryonic and Fetal Development; D005260:Female; D006041:Goats / Q000235:genetics* / Q000502:physiology; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008892:Milk / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D011247:Pregnancy; D011994:Recombinant Proteins / Q000378:metabolism; D012098:Reproduction
D011994:Recombinant Proteins; D000990:Antithrombin III