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Targeting a pre-mRNA structure with bipartite antisense molecules modulates tau alternative splicing.
Nucleic Acids Res 2012; 40(19):9836-49NA

Abstract

Approximately 15% of human genetic diseases are estimated to involve dysregulation of alternative pre-mRNA splicing. Antisense molecules designed to alter these and other splicing events typically target continuous linear sequences of the message. Here, we show that a structural feature in a pre-mRNA can be targeted by bipartite antisense molecules designed to hybridize with the discontinuous elements that flank the structure and thereby alter splicing. We targeted a hairpin structure at the boundary between exon 10 and intron 10 of the pre-mRNA of tau. Mutations in this region that are associated with certain forms of frontotemporal dementia, destabilize the hairpin to cause increased inclusion of exon 10. Via electrophoretic mobility shift and RNase protection assays, we demonstrate that bipartite antisense molecules designed to simultaneously interact with the available sequences that immediately flank the tau pre-mRNA hairpin do indeed bind to this structured region. Moreover, these agents inhibit exon 10 splicing and reverse the effect of destabilizing disease-causing mutations, in both in vitro splicing assays and cell culture. This general bipartite antisense strategy could be employed to modulate other splicing events that are regulated by RNA secondary structure.

Authors+Show Affiliations

Center for Neurologic Diseases, Harvard Medical School and Brigham and Women's Hospital, 77 Avenue Louis Pasteur, HIM 754, Boston, MA 02115, USA.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

22844088

Citation

Peacey, Eleanor, et al. "Targeting a pre-mRNA Structure With Bipartite Antisense Molecules Modulates Tau Alternative Splicing." Nucleic Acids Research, vol. 40, no. 19, 2012, pp. 9836-49.
Peacey E, Rodriguez L, Liu Y, et al. Targeting a pre-mRNA structure with bipartite antisense molecules modulates tau alternative splicing. Nucleic Acids Res. 2012;40(19):9836-49.
Peacey, E., Rodriguez, L., Liu, Y., & Wolfe, M. S. (2012). Targeting a pre-mRNA structure with bipartite antisense molecules modulates tau alternative splicing. Nucleic Acids Research, 40(19), pp. 9836-49. doi:10.1093/nar/gks710.
Peacey E, et al. Targeting a pre-mRNA Structure With Bipartite Antisense Molecules Modulates Tau Alternative Splicing. Nucleic Acids Res. 2012;40(19):9836-49. PubMed PMID: 22844088.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Targeting a pre-mRNA structure with bipartite antisense molecules modulates tau alternative splicing. AU - Peacey,Eleanor, AU - Rodriguez,Lilia, AU - Liu,Yang, AU - Wolfe,Michael S, Y1 - 2012/07/25/ PY - 2012/7/31/entrez PY - 2012/7/31/pubmed PY - 2013/1/15/medline SP - 9836 EP - 49 JF - Nucleic acids research JO - Nucleic Acids Res. VL - 40 IS - 19 N2 - Approximately 15% of human genetic diseases are estimated to involve dysregulation of alternative pre-mRNA splicing. Antisense molecules designed to alter these and other splicing events typically target continuous linear sequences of the message. Here, we show that a structural feature in a pre-mRNA can be targeted by bipartite antisense molecules designed to hybridize with the discontinuous elements that flank the structure and thereby alter splicing. We targeted a hairpin structure at the boundary between exon 10 and intron 10 of the pre-mRNA of tau. Mutations in this region that are associated with certain forms of frontotemporal dementia, destabilize the hairpin to cause increased inclusion of exon 10. Via electrophoretic mobility shift and RNase protection assays, we demonstrate that bipartite antisense molecules designed to simultaneously interact with the available sequences that immediately flank the tau pre-mRNA hairpin do indeed bind to this structured region. Moreover, these agents inhibit exon 10 splicing and reverse the effect of destabilizing disease-causing mutations, in both in vitro splicing assays and cell culture. This general bipartite antisense strategy could be employed to modulate other splicing events that are regulated by RNA secondary structure. SN - 1362-4962 UR - https://www.unboundmedicine.com/medline/citation/22844088/Targeting_a_pre_mRNA_structure_with_bipartite_antisense_molecules_modulates_tau_alternative_splicing_ L2 - https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gks710 DB - PRIME DP - Unbound Medicine ER -