Fig. 1

CRISPR/Cas9 generated chd7 mutants recapitulate CHARGE syndrome related phenotypes. (A) Exon‐intron structure of zebrafish chd7 with CRISPR exon 9 and exon 16 targeted regions noted; amino acid alignment and conservation of Exon 9, 2nd chromodomain target across species, identified 7 bp deletion from mutagenesis; amino acid alignment of Exon 16, ATP‐helicase domain target, and identified 1 bp deletion from mutagenesis; predicted Chd7 protein domains in wild‐type (top), ATP‐helicase domain 1 bp deletion allele (chd7 ^{rdu1002/rdu1002} ), and chromodomain 7 bp deletion allele (chd7 ^{ncu101/ncu101} ), additional target regions from sgRNAs noted. (B) Survival rate (%) of chd7 ^+/+ (n = 19), chd7 ^ncu101/+ (n = 54), and chd7 ^{ncu101/ncu101} (n = 25) siblings during a 30‐day period. (C) qPCR analysis of chd7 mRNA in 5 dpf larvae (chd7 ^ncu101/, chromodomain allele), normalized to wild‐type siblings. Each point represents a biological replicate of pooled larvae (per pool, chd7 ^+/+ n ~ 12–13; chd7 ^ncu101/+ n ~ 20–21; chd7 ^{ncu101/ncu101} n ~ 13–14). (D) qPCR analysis of chd7 mRNA in 5 dpf larvae (chd7 ^rdu1002/, ATP‐helicase domain allele), normalized to wild‐type siblings. Each point represents a biological replicate of pooled larvae (per pool, chd7 ^+/+ n ~ 13–14; chd7 ^rdu1002/− n ~ 22–23; chd7 ^{rdu1002/rdu1002} n ~ 11–12) (mean ± SD, Ordinary one‐way ANOVA with Tukey's multiple comparisons, *p < 0.05, **p < 0.01, ***p < 0.001,****p < 0.0001). (E) Bright‐field images of 5 dpf larvae (chd7 ^+/+, chd7 ^ncu101/+, chd7 ^{ncu101/ncu101} ) with examples of morphological phenotypes including uninflated swim bladder (asterisk), pericardial edema (triangle), and craniofacial defects (solid triangle). (F) Examples of varying otolith defects in chd7 ^ncu101/, dashed orange lines represent morphology of the otic vesicle, and anterior and posterior otoliths.