ddx52 is the pan gene. (A) High-resolution mapping of linked SNP markers in pan. Six and 16 recombinants were respectively identified for SNP37688707 and SNP37971903. SNP38867230 in ddx52 exon 14 was used for detection of the pan allele. (B) Sanger sequencing readouts from pan cDNA. The ddx52 sequence contains an in-frame indel that affects exons 13 and 14. (C) The three major structural domains of ddx52: Q motif, DEAD-box domain and helicase C domain. Gray color and scribble lines indicate regions that are affected in the pan allele and the ddx52del allele. (D) Illustration of and Sanger sequencing readouts depicting the sgRNA targeting site. The resulting ddx52del allele produces an out-of-frame indel that leads to an early stop codon in exon 2. (E) The ddx52del allele fails to complement the pan tailfin regeneration defect in adult animals. All adult animals with or without regeneration defects at the restrictive temperature had expected genotypes (n=23 pan/+, 12 pan/ddx52del). Red arrows indicate the amputation plane. Scale bar: 1 mm. (F) Whole-mount images of ddx52del×ddx52del embryos at 26°C. Approximately 25% of embryos had severe developmental defects at 6 days post-fertilization (dpf). Scale bar: 0.5 mm.
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