Figure 7

Phenotypic and genotypic analysis of embryos from an incross of hmx3a^SU42/+ parents injected with hmx2^MENTHU CRISPR reagents. (A and B) Lateral views of ear phenotypes at 4 day in live uninjected (A) and hmx2^MENTHU CRISPR-injected (B) hmx3a^SU42 homozygous mutant embryos. Rostral, left; Dorsal, top. The uninjected hmx3a^SU42 homozygous mutant embryo has two normal otoliths in each ear (A). In contrast, the hmx2^MENTHU CRISPR-injected hmx3a^SU42 homozygous mutant embryo has fused otoliths in both ears. (C and D) Wild-type (top row) and hmx2^MENTHU (bottom row) genomic sequences on top. Each colored box represents a specific nucleotide in the hmx2 coding sequence: A, green; C, blue; G, black; T, red. The hmx2^MENTHU mutant sequence contains a 5 bp deletion (CGCAG, red line), which introduces a premature stop codon (red dashed box) 14–16 bases after the deletion. Sequencing traces (below) from individual embryos from an incross of hmx3a^SU42/+ parents injected with hmx2^MENTHU CRISPR reagents. In the injected embryos, hmx2^MENTHU mutant sequences (with the 5 bp deletion) comprise ∼60% (C) to 90% (D) of all amplified sequences at this locus. Bar, 50 µm (A and B).