Abstract
Numerous genetic disorders are caused by loss-of-function mutations that disrupt the open reading frame of the gene either by nonsense or by frameshift (insertion, deletion, indel, or splicing) mutations. Most of the time, the result is the absence of functional protein synthesis due to mRNA degradation by nonsense-mediated mRNA decay, or rapid degradation of a truncated protein. Antisense-based splicing modulation is a powerful tool that has the potential to treat genetic disorders by restoring the open reading frame through selective removal of the mutated exon, or by restoring correct splicing.
We have developed this approach for a severe genetic skin disorder, recessive dystrophic epidermolysis bullosa, caused by mutations in the COL7A1 gene encoding type VII collagen. This gene is particularly suited for exon-skipping approaches due to its unique genomic structure. It is composed of 118 exons, 83 of which are in frame. Moreover, these exons encode a single repetitive collagenous domain.
Using this gene as an example, we describe general methods that demonstrate the feasibility and efficacy of the antisense-mediated exon-skipping strategy to reframe transcripts.
Sandrina Turczynski and Matthias Titeux have contributed equally to this work.
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Acknowledgments
The authors would like to thank Laure Tonasso and Audrey Decha for their help during the first stages of this project, as well as Dr Olivier Danos and Dr Luis Garcia for fruitful discussions. This work was supported by AFM (Association Française contre les Myopathies), EBAE (Epidermolyse Bulleuse Association d’Entraide), and ANR (Agence Nationale pour la Recherche, programme blanc 2007).
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Turczynski, S., Titeux, M., Pironon, N., Hovnanian, A. (2012). Antisense-Mediated Exon Skipping to Reframe Transcripts. In: Aartsma-Rus, A. (eds) Exon Skipping. Methods in Molecular Biology, vol 867. Humana Press. https://doi.org/10.1007/978-1-61779-767-5_15
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DOI: https://doi.org/10.1007/978-1-61779-767-5_15
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