Targeted disruption of the alpha1,3-galactosyltransferase gene in cloned pigs

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 f...

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Bibliographic Details
Published in:Nature biotechnology 2002-03, Vol.20 (3), p.251
Main Authors: Dai, Yifan, Vaught, Todd D, Boone, Jeremy, Chen, Shu-Hung, Phelps, Carol J, Ball, Suyapa, Monahan, Jeff A, Jobst, Peter M, McCreath, Kenneth J, Lamborn, Ashley E, Cowell-Lucero, Jamie L, Wells, Kevin D, Colman, Alan, Polejaeva, Irina A, Ayares, David L
Format: Article
Language:English
Subjects:
Animals
Blotting, Southern
Cell Line
Cell Nucleus - metabolism
Cloning, Organism
Epitopes
Female
Fibroblasts - metabolism
Galactosyltransferases - genetics
Male
Models, Genetic
Mutagenesis, Site-Directed
Polymerase Chain Reaction
Recombination, Genetic
Swine - genetics
Transfection
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Summary: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.
ISSN:1087-0156