title: Targeted disruption of the alpha1,3-galactosyltransferase gene in cloned pigs.
abstract: Galactose-alpha1,3-galactose (alpha1,3Gal) is the major xenoantigen causing hyperacute rejection in pig-to-human xenotransplantation. Disruption of the gene encoding pig alpha1,3-galactosyltransferase (alpha1,3GT) by homologous recombination is a means to completely remove the alpha1,3Gal epitopes from xenografts. Here we report the disruption of one allele of the pig alpha1,3GT gene in both male and female porcine primary fetal fibroblasts. Targeting was confirmed in 17 colonies by Southern blot analysis, and 7 of them were used for nuclear transfer. Using cells from one colony, we produced six cloned female piglets, of which five were of normal weight and apparently healthy. Southern blot analysis confirmed that these five piglets contain one disrupted pig alpha1,3GT allele.
authors.affiliation: PPL Therapeutics Inc., 1700 Kraft Drive, Blacksburg, VA 24060, USA. ydai@ppl-therapeutics.com
authors.forename: Yifan
authors.initials: Y
authors.lastname: Dai
authors.forename: Todd D
authors.initials: TD
authors.lastname: Vaught
authors.forename: Jeremy
authors.initials: J
authors.lastname: Boone
authors.forename: Shu-Hung
authors.initials: SH
authors.lastname: Chen
authors.forename: Carol J
authors.initials: CJ
authors.lastname: Phelps
authors.forename: Suyapa
authors.initials: S
authors.lastname: Ball
authors.forename: Jeff A
authors.initials: JA
authors.lastname: Monahan
authors.forename: Peter M
authors.initials: PM
authors.lastname: Jobst
authors.forename: Kenneth J
authors.initials: KJ
authors.lastname: McCreath
authors.forename: Ashley E
authors.initials: AE
authors.lastname: Lamborn
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Cowell-Lucero
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Wells
authors.forename: Alan
authors.initials: A
authors.lastname: Colman
authors.forename: Irina A
authors.initials: IA
authors.lastname: Polejaeva
authors.forename: David L
authors.initials: DL
authors.lastname: Ayares
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D019976:Cloning, Organism; D000939:Epitopes; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D005700:Galactosyltransferases / Q000235:genetics*; D008297:Male; D008957:Models, Genetic; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D013552:Swine / Q000235:genetics*; D014162:Transfection
chemical_list: D000939:Epitopes; D005700:Galactosyltransferases; C047006:N-acetyllactosaminide alpha-1,3-galactosyltransferase