Gene targeting in primary fetal fibroblasts from sheep and pig.
Nuclear transfer offers a new cell-based route for introducing precise genetic modifications in a range of animal species. However, significant challenges, such as establishment of somatic gene targeting techniques, must be overcome before the technology can be applied routinely. In this report, we describe targeted deletion at the GGTA1 (alpha 1,3-galactosyl transferase) and PrP (prion protein) loci in primary fibroblasts from livestock. We place particular emphasis on the growth characteristics of the primary cell cultures, since these are key to determining success.
Denning
C
C
Roslin Institute, Roslin, Midlothian, United Kingdom.
Dickinson
P
P
Burl
S
S
Wylie
D
D
Fletcher
J
J
Clark
A J
AJ
D000818:Animals; D000831:Animals, Newborn / Q000235:genetics*; D015152:Blotting, Northern; D002467:Cell Nucleus / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D005333:Fetus / Q000166:cytology*; D005347:Fibroblasts / Q000378:metabolism; D005700:Galactosyltransferases / Q000235:genetics*; D017353:Gene Deletion; D018390:Gene Targeting / Q000379:methods*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D007621:Karyotyping; D008957:Models, Genetic; D053652:Nuclear Transfer Techniques; D016133:Polymerase Chain Reaction; D011328:Prions / Q000235:genetics*; D012756:Sheep; D013552:Swine
D017931:DNA Primers; D011328:Prions; D005700:Galactosyltransferases; C047006:N-acetyllactosaminide alpha-1,3-galactosyltransferase