Figure 1

Generation of the samhd1 ^Δ23/Δ23 line. (A) gRNA sequence (red) designed to target a site within exon 4 of the samhd1 gene to initiate a 23 bp deletion in injected embryos. (B) Sanger sequencing of WT and samhd1 ^Δ23/Δ23 mutants from fish derived from F1 generation incross. Green highlighted area on WT sequencing represents the site of the 23 bp deletion, causing a frameshift on the mutant sequence. (C) Amplification of samhd1 transcripts from mutant and WT zebrafish embryos using exon 4 specific primers, yielded a single PCR product with clearly visible reduction in band size, as a result of 23 bp deletion. Length of samhd1 exon 4 is 161 bp in WT embryos, with the mutants producing a band of 138 bp. (D) qPCR analysis of samhd1 gene expression in 5 dpf WT and samhd1 ^Δ23/Δ23 larvae. Data analyzed using Mann Whitney U test and presented as median ± IQR (*P=0.0286). n=20 larvae per group, per biological replicate, repeated 4 times.