title: Production of alpha 1,3-galactosyltransferase-deficient pigs.
abstract: The enzyme alpha1,3-galactosyltransferase (alpha1,3GT or GGTA1) synthesizes alpha1,3-galactose (alpha1,3Gal) epitopes (Galalpha1,3Galbeta1,4GlcNAc-R), which are the major xenoantigens causing hyperacute rejection in pig-to-human xenotransplantation. Complete removal of alpha1,3Gal from pig organs is the critical step toward the success of xenotransplantation. We reported earlier the targeted disruption of one allele of the alpha1,3GT gene in cloned pigs. A selection procedure based on a bacterial toxin was used to select for cells in which the second allele of the gene was knocked out. Sequencing analysis demonstrated that knockout of the second allele of the alpha1,3GT gene was caused by a T-to-G single point mutation at the second base of exon 9, which resulted in inactivation of the alpha1,3GT protein. Four healthy alpha1,3GT double-knockout female piglets were produced by three consecutive rounds of cloning. The piglets carrying a point mutation in the alpha1,3GT gene hold significant value, as they would allow production of alpha1,3Gal-deficient pigs free of antibiotic-resistance genes and thus have the potential to make a safer product for human use.
authors.affiliation: PPL Therapeutics Inc., 1700 Kraft Drive, Blacksburg, VA 24060, USA.
authors.forename: Carol J
authors.initials: CJ
authors.lastname: Phelps
authors.forename: Chihiro
authors.initials: C
authors.lastname: Koike
authors.forename: Todd D
authors.initials: TD
authors.lastname: Vaught
authors.forename: Jeremy
authors.initials: J
authors.lastname: Boone
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Wells
authors.forename: Shu-Hung
authors.initials: SH
authors.lastname: Chen
authors.forename: Suyapa
authors.initials: S
authors.lastname: Ball
authors.forename: Susan M
authors.initials: SM
authors.lastname: Specht
authors.forename: Irina A
authors.initials: IA
authors.lastname: Polejaeva
authors.forename: Jeff A
authors.initials: JA
authors.lastname: Monahan
authors.forename: Pete M
authors.initials: PM
authors.lastname: Jobst
authors.forename: Sugandha B
authors.initials: SB
authors.lastname: Sharma
authors.forename: Ashley E
authors.initials: AE
authors.lastname: Lamborn
authors.forename: Amy S
authors.initials: AS
authors.lastname: Garst
authors.forename: Marilyn
authors.initials: M
authors.lastname: Moore
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Demetris
authors.forename: William A
authors.initials: WA
authors.lastname: Rudert
authors.forename: Rita
authors.initials: R
authors.lastname: Bottino
authors.forename: Suzanne
authors.initials: S
authors.lastname: Bertera
authors.forename: Massimo
authors.initials: M
authors.lastname: Trucco
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Starzl
authors.forename: Yifan
authors.initials: Y
authors.lastname: Dai
authors.forename: David L
authors.initials: DL
authors.lastname: Ayares
mesh_terms: D000483:Alleles; D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology; D002460:Cell Line; D003001:Cloning, Molecular; D019976:Cloning, Organism; D018076:DNA, Complementary; D004624:Embryo Transfer; D004768:Enterotoxins / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts; D005700:Galactosyltransferases / Q000172:deficiency* / Q000235:genetics*; D018390:Gene Targeting; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood; D016381:Islets of Langerhans Transplantation; D051379:Mice; D018345:Mice, Knockout; D017354:Point Mutation; D011247:Pregnancy; D013552:Swine / Q000235:genetics*; D014162:Transfection; D014183:Transplantation, Heterologous; D014312:Trisaccharides / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology
chemical_list: D001427:Bacterial Toxins; D018076:DNA, Complementary; D004768:Enterotoxins; D007075:Immunoglobulin M; D014312:Trisaccharides; C076569:alpha-galactosyl epitope; C041348:tcdA protein, Clostridium difficile; D005700:Galactosyltransferases; C047006:N-acetyllactosaminide alpha-1,3-galactosyltransferase