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Target selection for antisense oligonucleotide induced exon skipping in the dystrophin gene.
J Gene Med. 2003 Jun; 5(6):518-27.JG

Abstract

BACKGROUND

Duchenne muscular dystrophy (DMD) is an X-linked recessive muscle wasting disorder characterised by the absence of the protein dystrophin. Antisense oligonucleotides have been used to re-direct dystrophin pre-mRNA processing by blocking sequences crucial to pre-mRNA splicing, thereby inducing skipping of specific exons. We wished to determine which splicing motifs are most amenable as targets for antisense oligonucleotide induction of efficient and specific skipping of selected exons.

METHODS

Antisense oligonucleotides were directed at regions of dystrophin exon 19 involved in pre-mRNA splicing, including the donor and acceptor splice sites and the exon splicing enhancer (ESE). Cultured myotubes were transfected with antisense oligonucleotides at various concentrations and studies undertaken to determine both specificity and efficiency of induced exon 19 skipping.

RESULTS

Antisense oligonucleotides as small as 12 nucleotides targeting the ESE induced consistent and specific skipping of only exon 19 in both human and normal and mdx mouse myotubes. Antisense oligonucleotides directed at the donor and acceptor splice sites also induced specific exon 19 skipping while mismatched antisense oligonucleotides could only induce skipping when delivered at higher concentrations. No other dystrophin exons were removed from the mature mRNA as a consequence of these antisense oligonucleotides treatments.

CONCLUSIONS

Antisense oligonucleotides directed at the ESE tended to be marginally more efficient than those which targeted the donor or acceptor splice sites, based on their ability to induce specific skipping at lower concentrations. The specificity of exon removal does not appear to be a function of target selection, but may reflect the combination of the splicing motifs and position of that exon in the pre-mRNA.

Authors+Show Affiliations

Edith Cowan University, Perth, Western Australia, 6027.No 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

12797117

Citation

Errington, Stephen J., et al. "Target Selection for Antisense Oligonucleotide Induced Exon Skipping in the Dystrophin Gene." The Journal of Gene Medicine, vol. 5, no. 6, 2003, pp. 518-27.
Errington SJ, Mann CJ, Fletcher S, et al. Target selection for antisense oligonucleotide induced exon skipping in the dystrophin gene. J Gene Med. 2003;5(6):518-27.
Errington, S. J., Mann, C. J., Fletcher, S., & Wilton, S. D. (2003). Target selection for antisense oligonucleotide induced exon skipping in the dystrophin gene. The Journal of Gene Medicine, 5(6), 518-27.
Errington SJ, et al. Target Selection for Antisense Oligonucleotide Induced Exon Skipping in the Dystrophin Gene. J Gene Med. 2003;5(6):518-27. PubMed PMID: 12797117.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Target selection for antisense oligonucleotide induced exon skipping in the dystrophin gene. AU - Errington,Stephen J, AU - Mann,Christopher J, AU - Fletcher,Sue, AU - Wilton,Stephen D, PY - 2003/6/11/pubmed PY - 2004/2/18/medline PY - 2003/6/11/entrez SP - 518 EP - 27 JF - The journal of gene medicine JO - J Gene Med VL - 5 IS - 6 N2 - BACKGROUND: Duchenne muscular dystrophy (DMD) is an X-linked recessive muscle wasting disorder characterised by the absence of the protein dystrophin. Antisense oligonucleotides have been used to re-direct dystrophin pre-mRNA processing by blocking sequences crucial to pre-mRNA splicing, thereby inducing skipping of specific exons. We wished to determine which splicing motifs are most amenable as targets for antisense oligonucleotide induction of efficient and specific skipping of selected exons. METHODS: Antisense oligonucleotides were directed at regions of dystrophin exon 19 involved in pre-mRNA splicing, including the donor and acceptor splice sites and the exon splicing enhancer (ESE). Cultured myotubes were transfected with antisense oligonucleotides at various concentrations and studies undertaken to determine both specificity and efficiency of induced exon 19 skipping. RESULTS: Antisense oligonucleotides as small as 12 nucleotides targeting the ESE induced consistent and specific skipping of only exon 19 in both human and normal and mdx mouse myotubes. Antisense oligonucleotides directed at the donor and acceptor splice sites also induced specific exon 19 skipping while mismatched antisense oligonucleotides could only induce skipping when delivered at higher concentrations. No other dystrophin exons were removed from the mature mRNA as a consequence of these antisense oligonucleotides treatments. CONCLUSIONS: Antisense oligonucleotides directed at the ESE tended to be marginally more efficient than those which targeted the donor or acceptor splice sites, based on their ability to induce specific skipping at lower concentrations. The specificity of exon removal does not appear to be a function of target selection, but may reflect the combination of the splicing motifs and position of that exon in the pre-mRNA. SN - 1099-498X UR - https://www.unboundmedicine.com/medline/citation/12797117/Target_selection_for_antisense_oligonucleotide_induced_exon_skipping_in_the_dystrophin_gene_ L2 - https://doi.org/10.1002/jgm.361 DB - PRIME DP - Unbound Medicine ER -