Fig. 4

Zebrafish model of wars1 disruption shows microcephaly and supports a loss of function effect of the p.Arg133Cys variant. (a) Zebrafish wars1 knockouts generated by CRISPR/Cas9 technology and analysis of phenotypes in the F0 generation. Uninjected control (left panel), wars1 F0 mutant (middle panel), and wars1 F0 mutant rescue with wild-type human WARS1 mRNA (right panel) embryo at 3 dpf. Blue line indicates brain size. Red line indicates eye size. Black star indicates heart edema. (b) Quantification of head size from uninjected controls, wars1 F0, wars1 F0, and wild-type WARS1 mRNA (+WARS1), and p.Arg133Cys variant WARS1 mRNA. (c) Quantification of eye size from uninjected controls, wars1 F0, wars1 F0, and wild-type WARS1 mRNA (+WARS1), and wars1 F0 and p.Arg133Cys variant WARS1 mRNA. Each group represents ~30 embryos, and each dot shows a single embryo. Error bar = mean ± SD. For statistics, one-way analysis of variance (ANOVA) with Tukey's multiple comparisons test was used: ns, not significant p ≥ .05, **p < .01, ***p < .001, and ****p < .0001. All marked groups were compared to wars1 F0 mutant embryos. mRNA, messenger RNA.