Here, we report the identification and cDNA analysis of four intronic and three exonic OPA1 gene mutations that cause a variety of splicing defects including activation of cryptic splice sites in either flanking exon or intron sequences, and a leaky splicing mutation. Our results show that cDNA analysis is of prime importance for the full evaluation of the effect of putative splicing mutations in the OPA1 gene.
Article is a PDF, so annotating here.
Case#: OAK 96
DiseaseAssertion: clinical dx of adOA based on ophthalmological examination
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GenotypingMethod: All 30 coding exons of the OPA1 gene were amplified from genomic DNA by means of PCR. PCR and RT-PCR products were sequenced employing BigDyeTerminator Chemistry (Applied Biosystems, Darmstadt, Germany) and separated on an ABI 3100 DNA Sequencer
PreviouslyPublished: n/a
Variant: c.1705+1G>T (p.V556fsX596)
CAID: CA10576007
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AdditionalInfo: Generation of minigene constructs and heterologous expression in COS7 cells allowed for cDNA analysis of the c.1705+1G>T mutation. This revealed a deletion of the last 40 nucleotides in exon 17 resulting from a splicing event that involves a cryptic splice donor site within exon 17 resulting in a frameshift and premature translation termination (p.V556fsX596).