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Activation of a cryptic 5' splice site reverses the impact of pathogenic splice site mutations in the spinal muscular atrophy gene.
Nucleic Acids Res. 2017 Dec 01; 45(21):12214-12240.NA

Abstract

Spinal muscular atrophy (SMA) is caused by deletions or mutations of the Survival Motor Neuron 1 (SMN1) gene coupled with predominant skipping of SMN2 exon 7. The only approved SMA treatment is an antisense oligonucleotide that targets the intronic splicing silencer N1 (ISS-N1), located downstream of the 5' splice site (5'ss) of exon 7. Here, we describe a novel approach to exon 7 splicing modulation through activation of a cryptic 5'ss (Cr1). We discovered the activation of Cr1 in transcripts derived from SMN1 that carries a pathogenic G-to-C mutation at the first position (G1C) of intron 7. We show that Cr1-activating engineered U1 snRNAs (eU1s) have the unique ability to reprogram pre-mRNA splicing and restore exon 7 inclusion in SMN1 carrying a broad spectrum of pathogenic mutations at both the 3'ss and 5'ss of the exon 7. Employing a splicing-coupled translation reporter, we demonstrate that mRNAs generated by an eU1-induced activation of Cr1 produce full-length SMN. Our findings underscore a wider role for U1 snRNP in splicing regulation and reveal a novel approach for the restoration of SMN exon 7 inclusion for a potential therapy of SMA.

Authors+Show Affiliations

Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA.Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA. Interdepartmental Genetics and Genomics Program, Iowa State University, Ames, IA 50011, USA.Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA.Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA.Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA.Department of Biomedical Sciences, Iowa State University, Ames, IA 50011, USA. Interdepartmental Genetics and Genomics Program, Iowa State University, Ames, IA 50011, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

28981879

Citation

Singh, Natalia N., et al. "Activation of a Cryptic 5' Splice Site Reverses the Impact of Pathogenic Splice Site Mutations in the Spinal Muscular Atrophy Gene." Nucleic Acids Research, vol. 45, no. 21, 2017, pp. 12214-12240.
Singh NN, Del Rio-Malewski JB, Luo D, et al. Activation of a cryptic 5' splice site reverses the impact of pathogenic splice site mutations in the spinal muscular atrophy gene. Nucleic Acids Res. 2017;45(21):12214-12240.
Singh, N. N., Del Rio-Malewski, J. B., Luo, D., Ottesen, E. W., Howell, M. D., & Singh, R. N. (2017). Activation of a cryptic 5' splice site reverses the impact of pathogenic splice site mutations in the spinal muscular atrophy gene. Nucleic Acids Research, 45(21), 12214-12240. https://doi.org/10.1093/nar/gkx824
Singh NN, et al. Activation of a Cryptic 5' Splice Site Reverses the Impact of Pathogenic Splice Site Mutations in the Spinal Muscular Atrophy Gene. Nucleic Acids Res. 2017 Dec 1;45(21):12214-12240. PubMed PMID: 28981879.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Activation of a cryptic 5' splice site reverses the impact of pathogenic splice site mutations in the spinal muscular atrophy gene. AU - Singh,Natalia N, AU - Del Rio-Malewski,José Bruno, AU - Luo,Diou, AU - Ottesen,Eric W, AU - Howell,Matthew D, AU - Singh,Ravindra N, PY - 2017/05/29/received PY - 2017/09/06/accepted PY - 2017/10/6/pubmed PY - 2017/12/27/medline PY - 2017/10/6/entrez SP - 12214 EP - 12240 JF - Nucleic acids research JO - Nucleic Acids Res. VL - 45 IS - 21 N2 - Spinal muscular atrophy (SMA) is caused by deletions or mutations of the Survival Motor Neuron 1 (SMN1) gene coupled with predominant skipping of SMN2 exon 7. The only approved SMA treatment is an antisense oligonucleotide that targets the intronic splicing silencer N1 (ISS-N1), located downstream of the 5' splice site (5'ss) of exon 7. Here, we describe a novel approach to exon 7 splicing modulation through activation of a cryptic 5'ss (Cr1). We discovered the activation of Cr1 in transcripts derived from SMN1 that carries a pathogenic G-to-C mutation at the first position (G1C) of intron 7. We show that Cr1-activating engineered U1 snRNAs (eU1s) have the unique ability to reprogram pre-mRNA splicing and restore exon 7 inclusion in SMN1 carrying a broad spectrum of pathogenic mutations at both the 3'ss and 5'ss of the exon 7. Employing a splicing-coupled translation reporter, we demonstrate that mRNAs generated by an eU1-induced activation of Cr1 produce full-length SMN. Our findings underscore a wider role for U1 snRNP in splicing regulation and reveal a novel approach for the restoration of SMN exon 7 inclusion for a potential therapy of SMA. SN - 1362-4962 UR - https://www.unboundmedicine.com/medline/citation/28981879/Activation_of_a_cryptic_5'_splice_site_reverses_the_impact_of_pathogenic_splice_site_mutations_in_the_spinal_muscular_atrophy_gene_ L2 - https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gkx824 DB - PRIME DP - Unbound Medicine ER -