Correction of disease-associated exon skipping by synthetic exon-specific activators


Differential exon use is a hallmark of alternative splicing, a prevalent mechanism for generating protein isoform diversity. Many disease-associated mutations also affect pre-mRNA splicing, usually causing inappropriate exon skipping. SR proteins are essential splicing factors that recognize exonic splicing enhancers and drive exon inclusion. To emulate this function of SR proteins, we designed small chimeric effectors comprising a minimal synthetic RS domain covalently linked to an antisense moiety that targets an exon by Watson-Crick base pairing. Here we show that such synthetic effectors can mimic the functions of SR proteins and specifically restore wild type splicing when directed to defective BRCA1 or SMN2 pre-mRNA transcripts. This general approach can be used as a tool to investigate splicing mechanisms and modulate alternative splicing of specific genes, and as a therapeutic strategy to correct splicing defects responsible for numerous diseases.

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Figure 1: Design of synthetic compounds that specifically promote exon inclusion.
Figure 2: ESSENCE compounds restore BRCA1 E1694X splicing to wild type levels.
Figure 3: The RS minidomain is active in a phosphorylated state.
Figure 4: Rescue of SMN2 exon 7 splicing to SMN1 levels by ESSENCE.


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Supported by the NIH. We thank L. Manche for technical assistance, and R. Del Vecchio, M. Hastings and B. Stillman for helpful comments.

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Correspondence to Adrian R. Krainer.

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A patent application has been filed by the authors and Cold Spring Harbor Laboratory.

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Cartegni, L., Krainer, A. Correction of disease-associated exon skipping by synthetic exon-specific activators. Nat Struct Mol Biol 10, 120–125 (2003) doi:10.1038/nsb887

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