PCRs confirm expected CRISPR-Cas9 induced deletions in serpine3cbg17 and serpine3cbg18 fish.(A) Genotyping of the serpine3cbg17 with two primers (F0 generation). Injection of CRISPR guides into the one-cell embryo leads to mosaic embryos (several pooled at 72 hpf) with a strong wild type (WT) band at about 557 bp and a weak mutant band at about 161 bp, while uninjected embryos just have a single WT band (557 bp). The raw image is provided as Figure 4—source data 4. (B) Genotyping of serpine3cbg17 with a mix of three primers (F2 generation). Heterozygous animals (H) have two bands, a WT (expected height: 286 bp) and mutant band (expected height: 161 bp), while homozygous mutants (M) and homozygous WT animals show a single band. NC – negative control (water). The raw image is provided as Figure 4—source data 5. (C) Genotyping of serpine3cbg18 with two primers. Heterozygous animals (H) have two bands, a WT (expected height: 342 bp) and mutant band (expected height: 250 bp), while homozygous mutants (M) and homozygous WT animals show a single band. NC – negative control (water). The raw image is provided as Figure 4—source data 6.
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