Figure 4—figure supplement 1.

(A) 6 days post-fertilization (dpf) zebrafish embryos injected with either NT or tuba1a/c sg1 Alt-R CRISPR Cas9 gRNAs. (B) Validation of Alt-R CRISPR Cas9 tuba1a/c sg1 targeting with Sanger sequencing. Synthego Ice software estimated 88% indels in the tuba1a locus and 97% indels in the tuba1c locus. (C) Validation of Alt-R CRISPR Cas9 tuba1a-specific gRNA targeting with Sanger sequencing. Synthego Ice software estimated 97% indels in the tuba1a locus. (D) Validation of Alt-R CRISPR Cas9 tuba1c-specific gRNA targeting with Sanger sequencing. Synthego Ice software estimated 56% indels in the tuba1c locus. (E) Validation of Alt-R CRISPR Cas9 tuba1a/c sg2 targeting with Sanger sequencing. Synthego Ice software estimated 88% indels in the tuba1a locus and 86% indels in the tuba1c locus. (F) Percentage of NT or tuba1a/c sg2 Alt-R zebrafish embryos with dispersed melanocyte phenotypes. N=2 independent experiments. (G) Adult mitfa^Cas9 MG-tuba1a/c F0 fish. Image is representative of n=7 F0 fish. (H) Adult mitfa^Cas9 MG-tuba1a/c F1 fish. Image is representative of n=14 F1 fish. (I) CRISPR-seq results are shown for wild-type (WT) and n=2 F1 MG-tuba1a/c fish sorted for GFP+ cells. Results are shown as a fraction of sequences with indels calculated using CRISPResso.