Fig. 1

Generation of klotho gene knockout blastocysts by SCNT using non-selected porcine fibroblasts transfected with Cas9-sgRNA RNPs. a. Schematic representation of sgRNA specific to exon 3 of the porcine klotho locus. The sgRNA targeting sequence is highlighted in red and protospacer adjacent motif (PAM) is highlighted in blue. b. Comparison of preimplantation development of cloned embryos after SCNT with Cas9-sgRNA RNPs (targeting the klotho gene) transfected or non-transfected fibroblasts. Within each category, groups marked with different letters are significantly different (P < 0.05). c. Rate of DNA editing on the klotho gene of blastocysts generated by SCNT using non-selected porcine fibroblasts transfected with Cas9-sgRNA RNPs. d. T7 endonuclease I (T7E1) assay: the T7E1 assay was conducted using genomic DNA from 20 cloned blastocysts. (M, Marker; WT, wild-type; BL, blastocyst). For improved clarity and conciseness, cropped areas of the gel are shown. All cropped regions originate from the same gel (Additional file 2: Fig. S1). e. A diagram illustrating the editing scheme on exon 3 of the klotho gene of blastocysts