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Antisense oligonucleotide therapy for splicing defects in OPA 1‐related dominant optic atrophy
Author(s) -
Wissinger B.,
Synofzik M.,
Schöls L.,
Bonifert T.
Publication year - 2017
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2017.03173
Subject(s) - splice , rna splicing , exon , transfection , mutant , microbiology and biotechnology , oligonucleotide , point mutation , messenger rna , biology , mutation , exon skipping , splice site mutation , alternative splicing , cell culture , gene , genetics , rna
Summary Mutations in OPA 1 are the main cause of dominant optic atrophy ( DOA ) and have also been implicated in a variety of syndromic neuropathies such as DOA plus or Behr‐like syndrome. We have recently discovered a disease‐causing deep intronic mutation ( DIM ) in OPA 1 that induces a constitutive inclusion of a cryptic exon into OPA 1 transcripts. As a potential therapeutic strategy we sought to correct mis‐splicing of the mutant pre‐ mRNA by using antisense oligonucleotides targeting the cryptic acceptor splice site and the novel branch point created by the DIM , respectively. Transfection of patient‐derived primary dermal fibroblasts with AON s resulted in splice correction of the mutant pre‐ mRNA in a time and concentration dependent manner. Maximal rescue efficacy of up ~55% was obtained with the cryptic acceptor splice site targeting AON . Peak efficacy of splice correction was observed at 4 days after treatment. However, significant effects were still seen 14 days post‐transfection in the proliferating cell culture. Western blot analysis revealed increased amounts of OPA 1 protein with maximum amounts at ~3 days post‐treatment. In summary, we provide the first mutation‐specific rescue strategy for OPA 1 deficiency using synthetic AON s.