Figure 1.

Generation of transgenic fibroblasts and pigs. (A) Schematic drawing of the genome editing approach. A donor DNA encoding for the point mutation c.2223 C > G was generated. The point mutation results in an additional BslI restriction site. A double strand break was introduced in the porcine genomic DNA by TALENs and the point mutation was integrated by homology directed repair. Screening was accomplished by PCR and subsequent BslI restriction. The wildtype (WT) DNA generates a 205 bp and the genome edited (R723G) DNA a 138 bp fragment. (B–D) Agarose gel electrophoresis of BslI restriction fragments from PCR on DNA from transfected cell cultures (B), piglets cloned from cell culture H4 (C), and piglets from a subclone of cell culture H4 and the wildtype porcine fetal fibroblasts (PFF) as control (D).