title: Efficient generation of alpha(1,3) galactosyltransferase knockout porcine fetal fibroblasts for nuclear transfer.
abstract: Pigs are currently considered the most likely source of organs for human xenotransplantation because of anatomical and physiological similarities to humans, and the relative ease with which they can be bred in large numbers. A severe form of rejection known as hyperacute rejection has been the major barrier to the use of xenografts. Generating transgenic pigs for organ transplantation is likely to involve precise genetic manipulation to ablate the alpha(1,3) galactosyltransferase (galT) gene. In contrast to the mouse, homologous recombination in livestock species to ablate genes is hampered by the inability to isolate functional embryonic stem cells. However, nuclear transfer using genetically targeted cultured somatic cells provides an alternative means to producing pigs deficient for galT. In this study we successfully produced galT+/- somatic porcine fetal fibroblasts using two approaches; positive negative selection (PNS) using an isogenic targeting construct, and with a promoterless vector using non-isogenic DNA.
authors.affiliation: Reproduction and Biotechnology Unit, BresaGen Limited, Adelaide, South Australia, Australia. sharon.harrison@adelaide.edu.au
authors.forename: Sharon J
authors.initials: SJ
authors.lastname: Harrison
authors.forename: Angelo
authors.initials: A
authors.lastname: Guidolin
authors.forename: Renate
authors.initials: R
authors.lastname: Faast
authors.forename: Lesley A
authors.initials: LA
authors.lastname: Crocker
authors.forename: Chris
authors.initials: C
authors.lastname: Giannakis
authors.forename: Anthony J F
authors.initials: AJ
authors.lastname: D'Apice
authors.forename: Mark B
authors.initials: MB
authors.lastname: Nottle
authors.forename: Ian
authors.initials: I
authors.lastname: Lyons
mesh_terms: D000818:Animals; D002469:Cell Separation; D005333:Fetus; D005347:Fibroblasts; D005700:Galactosyltransferases / Q000235:genetics*; D017353:Gene Deletion; D018390:Gene Targeting / Q000379:methods; D016133:Polymerase Chain Reaction; D013552:Swine
chemical_list: D005700:Galactosyltransferases