Transgenic cattle resulting from biopsied embryos: expression of c-ski in a transgenic calf.
Producing transgenic cattle by microinjection of DNA into pronuclei has been inefficient and costly, in large part because of the cost of maintaining numerous nontransgenic pregnancies to term. We designed a system for early identification of transgenic embryos in which biopsies of embryos were assayed by polymerase chain reaction for presence of the transgene before embryo transfer. A total of 2555 embryos were microinjected with one of two DNA constructs. Of the 533 embryos biopsied, 112 were judged to be potentially transgenic and were transferred nonsurgically to recipients, resulting in production of 29 putative transgenic fetuses. One fetus and one calf (7% of offspring) were subsequently shown to be definitively transgenic. The calf was transgenic for a chicken c-ski cDNA, and several months after birth developed dramatic muscular hypertrophy followed by muscle degeneration. This phenotype was associated with expression of high levels of mRNA from the transgene.
Bowen
R A
RA
Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins 80523.
Reed
M L
ML
Schnieke
A
A
Seidel
G E
GE
Stacey
A
A
Thomas
W K
WK
Kajikawa
O
O
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002417:Cattle / Q000196:embryology / Q000235:genetics*; D002645:Chickens / Q000235:genetics; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D008297:Male; D008845:Microinjections; D008969:Molecular Sequence Data; D009132:Muscles / Q000033:anatomy & histology; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D012045:Regulatory Sequences, Nucleic Acid; D009053:Sarcoma Viruses, Murine / Q000235:genetics