Figure 1
From: Generation of Fel d 1 chain 2 genome-edited cats by CRISPR-Cas9 system
Generation and cloning of CH2 genome-edited cats using the CRISPR-Cas9 system and Sanger sequencing. (a) The CH2 genome-edited founder cat (Male, KS-M-001, Heavy). (b) The CH2 genome-edited founder cat (Female, KS-F-011, Haemi). (c) The CH2 genome-edited cat (F1, Male, CH2−/−, M-008, Alsik). (d) The CH2 genome-edited cloned cat (Male, CH2-/-, KS-M-004, Alsik C) from “Alsik”. (e) The modified target region in each CH2 genome-edited cat. (f) The sequencing chromatogram of the target region of the CH2 genome-edited founder cat (Heavy). (g) The sequencing chromatogram of the target region of the CH2 genome-edited founder cat (Haemi). (h) The sequencing chromatogram of the target region of the CH2 genome-edited cat (Alsik). (i) The sequencing chromatogram of the target region of the CH2 genome-edited cloned cat (Alsik C). “Heavy” and “Haemi” were generated by cytoplasmic microinjection of sgRNA C2-1 and Cas9 mRNA and embryo transfer. “Alsik” was generated from the mating of “Heavy” and “Haemi”. “Alsik C” was cloned from “Alsik” via CICT. Through the use of PCR and Sanger sequencing, mutations are found in every CH2 genome-edited cat. The blue highlights indicate the Cas9-targeted CH2 sequences. The PAM sequence is highlighted in bold blue. Bold red is used to highlight the indels. Each CH2 genome-edited cat’s DNA sequence is in alignment with the WT cat’s DNA sequence. CICT; cytoplasm injection cloning technology. ID; identification. CH2; Fel d 1 chain 2. Indel; insertion/deletion. PAM; protospacer adjacent motif. WT; wild type. Ns; nucleotides. T; thymine. G; guanine.
