title: Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts.
abstract: Ovine primary fetal fibroblasts were cotransfected with a neomycin resistance marker gene (neo) and a human coagulation factor IX genomic construct designed for expression of the encoded protein in sheep milk. Two cloned transfectants and a population of neomycin (G418)-resistant cells were used as donors for nuclear transfer to enucleated oocytes. Six transgenic lambs were liveborn: Three produced from cloned cells contained factor IX and neo transgenes, whereas three produced from the uncloned population contained the marker gene only. Somatic cells can therefore be subjected to genetic manipulation in vitro and produce viable animals by nuclear transfer. Production of transgenic sheep by nuclear transfer requires fewer than half the animals needed for pronuclear microinjection.
authors.affiliation: PPL Therapeutics, Roslin, Midlothian, EH25 9PP, Scotland, UK.
authors.forename: A E
authors.initials: AE
authors.lastname: Schnieke
authors.forename: A J
authors.initials: AJ
authors.lastname: Kind
authors.forename: W A
authors.initials: WA
authors.lastname: Ritchie
authors.forename: K
authors.initials: K
authors.lastname: Mycock
authors.forename: A R
authors.initials: AR
authors.lastname: Scott
authors.forename: M
authors.initials: M
authors.lastname: Ritchie
authors.forename: I
authors.initials: I
authors.lastname: Wilmut
authors.forename: A
authors.initials: A
authors.lastname: Colman
authors.forename: K H
authors.initials: KH
authors.lastname: Campbell
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D019976:Cloning, Organism; D004351:Drug Resistance; D004624:Embryo Transfer; D005164:Factor IX / Q000096:biosynthesis / Q000235:genetics*; D005260:Female; D005333:Fetus; D005347:Fibroblasts; D005839:Gentamicins / Q000494:pharmacology; D006801:Humans; D008297:Male; D008892:Milk / Q000378:metabolism; D009355:Neomycin / Q000494:pharmacology; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology; D011994:Recombinant Proteins / Q000096:biosynthesis; D012756:Sheep / Q000196:embryology / Q000235:genetics*; D014162:Transfection; D019076:Transgenes
chemical_list: D005839:Gentamicins; D011994:Recombinant Proteins; D005164:Factor IX; C010680:antibiotic G 418; D009355:Neomycin