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Generation of a complete series of Irx cluster mutants in zebrafish. (A) Schematic of CRISPR/Cas9 strategy to generate Irx cluster mutants. All selected alleles result in frame-shift mutations (red arrowheads). Dashes indicate deleted nucleotides, and green letters indicate insertions. All alleles are predicted to delete or truncate the DNA-binding homeobox domain (blue box). (B) Brightfield images showing dorsal axis curvature in the absence of IrxA genes and cardiac edema in the absence of IrxB genes. (C) Summary of genotypes for the indicated phenotypes from a quintuple heterozygous in-cross. P-values report chi-square tests for deviation from the expected 1:2:1 wild type:heterozygous:homozygous Mendelian ratio. Scale bar: 1 mm.
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