Figure 2

Generation of zebrafish stag germline mutants. CRISPR-Cas9 genome editing was used to generate germline mutations in stag1a, stag1b, and stag2b. (A) Exon diagrams of the respective paralogues showing details of the editing strategy. sgRNA binding sites are marked by red arrowheads with the type of mutation generated indicated above. (B) mRNA levels of the four paralogues in each of the mutant lines indicated on the x-axis. Each data point represents mRNA isolated from a pool of 30 embryos. All graphs are mean +/- one standard deviation. *P ≤ 0.05, **P ≤ 0.01, and ****P ≤ 0.0001; one-way ANOVA. Expression was normalized to the reference gene, b-actin (Supplementary Figure 1C) (C)stag2b^nz207 mutants have displaced pigment cells in the tail fin, zoom-ins are shown in insets. Mutants also show mild developmental delay with late swim bladder inflation as indicated by the black arrow. Scale bars are 200 μm.