Skip to main content
Fig. 2 | BMC Genomics

Fig. 2

From: One-step generation of a targeted knock-in calf using the CRISPR-Cas9 system in bovine zygotes

Fig. 2

Development of a targeted knock-in bull calf. We monitored and analyzed the development of the SRY-GFP knock-in bull calf produced by cytoplasmic injection of a homology mediated end joining donor template and the CRISPR-Cas9 system in bovine zygotes. a ultrasound of the day 68 fetus revealing the male genital tubercle (arrow) caudal to the umbilicus indicating a male phenotype, b the SRY-GFP knock-in bull calf (Cosmo) at 2 days of age, c Analysis of SRY-GFP knock-in by the polymerase chain reaction (PCR). DNA was extracted from three tissue types: placental cotyledons (trophectodermal origin), blood and fibroblast cells (mesodermal origin). The donor plasmid was used as the positive control and water was used as the negative control. Expected band sizes: wild type 520 bp, SRY-GFP knock-in 3721 bp. The lower band from the calf runs higher than wild type due to the 26 bp insertion, and d Genotypic sex. Expected band sizes: female 208 bp; male 189 bp & 208 bp. lane 1 = wild type male; lane 2 = recipient female (3113); lane 3 = Cosmo placenta; lane 4 = Cosmo blood; lane 5 = Cosmo fibroblast; lane 6 = plasmid; lane 7 = water

Back to article page