Production of homozygous alpha-1,3-galactosyltransferase knockout pigs by breeding and somatic cell nuclear transfer.
We report here our experience regarding the production of double or homozygous Gal knockout (Gal KO) pigs by breeding and somatic cell nuclear transfer (SCNT). Large White x Landrace female heterozygous Gal KO founders produced using SCNT were mated with Hampshire or Duroc males to produce a F1 generation. F1 heterozygous pigs were then bred to half-sibs to produce a F2 generation which contained Gal KO pigs. To determine the viability of mating Gal KO pigs with each other, one female F2 Gal KO pig was bred to a half-sib and subsequently a full-sib Gal KO. F1 and F2 heterozygous females were also mated to F2 Gal KO males. All three types of matings produced Gal KO pigs. To produce Gal KO pigs by SCNT, heterozygous F1s were bred together and F2 fetuses were harvested to establish primary cultures of Gal KO fetal fibroblasts. Gal KO embryos were transferred to five recipients, one of which became pregnant and had a litter of four piglets. Together our results demonstrate that Gal KO pigs can be produced by breeding with each other and by SCNT using Gal KO fetal fibroblasts.
Research Centre for Reproductive Health & Discipline of Obstetrics and Gynaecology, School of Paediatrics and Reproductive Health, University of Adelaide, Adelaide, South Australia, Australia. mark.nottle@adelaide.edu.au
Mark B
MB
Nottle
Luke F S
LF
Beebe
Sharon J
SJ
Harrison
Stephen M
SM
McIlfatrick
Rodney J
RJ
Ashman
Phillip J
PJ
O'Connell
Evelyn J
EJ
Salvaris
Nella
N
Fisicaro
Sandra
S
Pommey
Peter J
PJ
Cowan
Anthony J F
AJ
d'Apice
D000822:Animal Husbandry / Q000379:methods; D000818:Animals; D030801:Animals, Genetically Modified; D017849:Animals, Inbred Strains / Q000276:immunology*; D005347:Fibroblasts; D005700:Galactosyltransferases / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D008297:Male; D009206:Myocardium / Q000276:immunology / Q000648:ultrastructure; D053652:Nuclear Transfer Techniques; D013552:Swine; D014183:Transplantation, Heterologous
D005700:Galactosyltransferases; C047006:N-acetyllactosaminide alpha-1,3-galactosyltransferase