Fig. 1

Optimization of the CRISPR/Cas9 system in sheep fibroblasts and embryos. A Schematic representation of the study design for the optimization of CRISPR/Cas9:sgRNA delivery system in fibroblasts and sheep microinjected zygotes. B Editing efficiency of different sgRNAs targeting the MSNT gene in sheep fibroblasts. C, D Embryo development rate and editing efficiency for each tested microinjection group. In groups #1, #2, and #3, Cas9 mRNA concentration was 25 ng/μL, whereas concentration of total sgRNAs was 100 ng/μL, 200 ng/μL, and 400 ng/μL, respectively. In groups #4, #5, and #6, Cas9 mRNA concentration was 50 ng/μL, while concentration of total sgRNAs was 100 ng/μL, 200 ng/μL, and 400 ng/μL, respectively. In groups #7, #8, and #9, Cas9 mRNA concentration was 100 ng/μL, while concentration of total sgRNAs was 100 ng/μL, 200 ng/μL, and 400 ng/μL, respectively. In groups #10, #11, and #12, Cas9 mRNA concentration was 400 ng/μL, while concentration of total sgRNAs was 100 ng/μL, 200 ng/μL, and 400 ng/μL, respectively