Journal article

An efficient method for producing alpha(1,3)-galactosyltransferase gene knockout pigs

S Harrison, A Boquest, C Grupen, R Faast, A Guildolin, C Giannakis, L Crocker, S McIlfatrick, R Ashman, J Wengle, I Lyons, P Tolstoshev, P Cowan, A Robins, P O'Connell, AJF D'Apice, M Nottle

CLONING AND STEM CELLS | MARY ANN LIEBERT, INC | Published : 2004

Abstract

We have reported relatively efficient methods for somatic cell nuclear transfer and for knocking out the alpha(1,3)-galactosyltransferase (alpha1,3-GT) gene in porcine fetal fibroblasts using a nonisogenic promoterless construct approach. Here we report the production of alpha1,3-GT gene knockout pigs using these procedures. Seven alpha1,3-GT gene knockout cell clones were identified by long-range PCR from 108 neomycin resistant (neo(R)) colonies, giving a 6.5% targeting efficiency. Three cell clones were used for nuclear transfer. Nuclear transfer was performed using a fusion before activation protocol using in vitro-matured adult oocytes. Between 51 and 110 fused couplets were transferred ..

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