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Fig. 2. Target sites of the dfna5 morpholinos and aberrant splicing of
dfna5 transcript. (A) Genomic organization of human DFNA5 and
zebrafish Dfna5. The exon/intron boundaries of DFNA5 are entirely
conserved between human and zebrafish. (a) Individuals with DFNA5
mutations carry an insertion/deletion mutation in intron 7, which leads to
skipping of exon 8 (indicated in red). Absence of exon 8 causes a frameshift
after amino acid 330, resulting in an aberrant stretch of 41 amino acids
followed by a premature stop. (b) The two morpholino antisense oligos designed
against Dfna5 mRNA are indicated in red. The splice site antisense
oligo ('GT-MO') directed against the donor site of exon 8 leads to skipping of
the targeted exon (indicated in red), resembling the human mutation. (B)
RT-PCR time course of aberrant splicing of dfna5 mRNA caused by the
dfna5 GT-MO. At 28 hpf, wild-type dfna5 transcript is not
detectable by RT-PCR, only PCR products lacking exon 8 (indicated with 
exon 8) or with partial retention of the following intron (arrowhead) are
present. Wild-type transcript recovers starting at day 2, but the
morpholino-modulated transcripts are still present at day 7.
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