Abstract
Exon skipping induced by gene mutations is a common mechanism responsible for many genetic diseases. A practical approach to correct the aberrant splicing of defective genes is to use antisense oligonucleotides (ASOs). The recognition of splice sites and the regulation of splicing involve multiple positive or negative cis-acting elements and trans-acting factors. Base-pairing of ASOs to a negative element in a targeted pre-mRNA blocks the binding of splicing repressors to this cis-element and/or disrupts an unfavorable secondary structure; as a result, the ASO restores exon inclusion. For example, we have recently shown that appropriate 2′-O-(2-methoxyethyl) (MOE) phosphorothioate-modified ASOs can efficiently correct survival motor neuron 2 (SMN2) exon 7 splicing in a cell-free splicing assay, in cultured human cells—including patient fibroblasts—and in both peripheral tissues and the CNS of SMA mouse models. These ASOs are promising drug leads for SMA therapy.
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Acknowledgments
The authors thank Drs C. Frank Bennett, Timothy A. Vickers, Brenda F. Baker, Gene Hung, and Frank Rigo at Isis Pharmaceuticals for providing MOE ASOs, for many helpful discussions, and sharing protocols, as well as Dr. Kentaro Sahashi at Cold Spring Harbor Laboratory for helping to implement the embryonic ICV injection procedure. The relevant research in the authors’ laboratory was generously supported by the Muscular Dystrophy Association, the National Institute of Neurological Disorders and Stroke, and the SMA Foundation.
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Hua, Y., Krainer, A.R. (2012). Antisense-Mediated Exon Inclusion. 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_20
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DOI: https://doi.org/10.1007/978-1-61779-767-5_20
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