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Antisense-induced exon skipping for duplications in Duchenne muscular dystrophy.
BMC Med Genet. 2007 Jul 05; 8:43.BM

Abstract

BACKGROUND

Antisense-mediated exon skipping is currently one of the most promising therapeutic approaches for Duchenne muscular dystrophy (DMD). Using antisense oligonucleotides (AONs) targeting specific exons the DMD reading frame is restored and partially functional dystrophins are produced. Following proof of concept in cultured muscle cells from patients with various deletions and point mutations, we now focus on single and multiple exon duplications. These mutations are in principle ideal targets for this approach since the specific skipping of duplicated exons would generate original, full-length transcripts.

METHODS

Cultured muscle cells from DMD patients carrying duplications were transfected with AONs targeting the duplicated exons, and the dystrophin RNA and protein were analyzed.

RESULTS

For two brothers with an exon 44 duplication, skipping was, even at suboptimal transfection conditions, so efficient that both exons 44 were skipped, thus generating, once more, an out-of-frame transcript. In such cases, one may resort to multi-exon skipping to restore the reading frame, as is shown here by inducing skipping of exon 43 and both exons 44. By contrast, in cells from a patient with an exon 45 duplication we were able to induce single exon 45 skipping, which allowed restoration of wild type dystrophin. The correction of a larger duplication (involving exons 52 to 62), by combinations of AONs targeting the outer exons, appeared problematic due to inefficient skipping and mistargeting of original instead of duplicated exons.

CONCLUSION

The correction of DMD duplications by exon skipping depends on the specific exons targeted. Its options vary from the ideal one, restoring for the first time the true, wild type dystrophin, to requiring more 'classical' skipping strategies, while the correction of multi-exon deletions may need the design of tailored approaches.

Authors+Show Affiliations

Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. a.m.rus@lumc.nlNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17612397

Citation

Aartsma-Rus, Annemieke, et al. "Antisense-induced Exon Skipping for Duplications in Duchenne Muscular Dystrophy." BMC Medical Genetics, vol. 8, 2007, p. 43.
Aartsma-Rus A, Janson AA, van Ommen GJ, et al. Antisense-induced exon skipping for duplications in Duchenne muscular dystrophy. BMC Med Genet. 2007;8:43.
Aartsma-Rus, A., Janson, A. A., van Ommen, G. J., & van Deutekom, J. C. (2007). Antisense-induced exon skipping for duplications in Duchenne muscular dystrophy. BMC Medical Genetics, 8, 43.
Aartsma-Rus A, et al. Antisense-induced Exon Skipping for Duplications in Duchenne Muscular Dystrophy. BMC Med Genet. 2007 Jul 5;8:43. PubMed PMID: 17612397.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Antisense-induced exon skipping for duplications in Duchenne muscular dystrophy. AU - Aartsma-Rus,Annemieke, AU - Janson,Anneke A M, AU - van Ommen,Gert-Jan B, AU - van Deutekom,Judith C T, Y1 - 2007/07/05/ PY - 2007/03/14/received PY - 2007/07/05/accepted PY - 2007/7/7/pubmed PY - 2007/7/28/medline PY - 2007/7/7/entrez SP - 43 EP - 43 JF - BMC medical genetics JO - BMC Med Genet VL - 8 N2 - BACKGROUND: Antisense-mediated exon skipping is currently one of the most promising therapeutic approaches for Duchenne muscular dystrophy (DMD). Using antisense oligonucleotides (AONs) targeting specific exons the DMD reading frame is restored and partially functional dystrophins are produced. Following proof of concept in cultured muscle cells from patients with various deletions and point mutations, we now focus on single and multiple exon duplications. These mutations are in principle ideal targets for this approach since the specific skipping of duplicated exons would generate original, full-length transcripts. METHODS: Cultured muscle cells from DMD patients carrying duplications were transfected with AONs targeting the duplicated exons, and the dystrophin RNA and protein were analyzed. RESULTS: For two brothers with an exon 44 duplication, skipping was, even at suboptimal transfection conditions, so efficient that both exons 44 were skipped, thus generating, once more, an out-of-frame transcript. In such cases, one may resort to multi-exon skipping to restore the reading frame, as is shown here by inducing skipping of exon 43 and both exons 44. By contrast, in cells from a patient with an exon 45 duplication we were able to induce single exon 45 skipping, which allowed restoration of wild type dystrophin. The correction of a larger duplication (involving exons 52 to 62), by combinations of AONs targeting the outer exons, appeared problematic due to inefficient skipping and mistargeting of original instead of duplicated exons. CONCLUSION: The correction of DMD duplications by exon skipping depends on the specific exons targeted. Its options vary from the ideal one, restoring for the first time the true, wild type dystrophin, to requiring more 'classical' skipping strategies, while the correction of multi-exon deletions may need the design of tailored approaches. SN - 1471-2350 UR - https://www.unboundmedicine.com/medline/citation/17612397/Antisense_induced_exon_skipping_for_duplications_in_Duchenne_muscular_dystrophy_ L2 - https://bmcmedgenet.biomedcentral.com/articles/10.1186/1471-2350-8-43 DB - PRIME DP - Unbound Medicine ER -