Correction of tau mis-splicing caused by FTDP-17 MAPT mutations by spliceosome-mediated RNA trans-splicing

Teresa Rodriguez-Martin, Karen Anthony, Mariano Garcia-Blanco, S. Gary Mansfield, Brian H. Anderton, Jean Marc Gallo

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17) is caused by mutations in the MAPT gene, encoding the tau protein that accumulates in intraneuronal lesions in a number of neurodegenerative diseases. Several FTDP-17 mutations affect alternative splicing and result in excess exon 10 (E10) inclusion in tau mRNA. RNA reprogramming using spliceosome-mediated RNA trans-splicing (SMaRT) could be a method of choice to correct aberrant E10 splicing resulting from FTDP-17 mutations. SMaRT creates a hybrid mRNA through a trans-splicing reaction between an endogenous target pre-mRNA and a pre- trans-splicing RNA molecule (PTM). However, FTDP-17 mutations affect the strength of cis-splicing elements and could favor cis-splicing over trans-splicing. Excess E10 inclusion in FTDP-17 can be caused by intronic mutations destabilizing a stem-loop protecting the 5′ splice site at the E10/intron 10 junction. COS cells transfected with a minigene containing the intronic +14 mutation produce exclusively E10+ RNA. Generation of E10- RNA was restored after co-transfection with a PTM designed to exclude E10. Similar results were obtained with a target containing the exonic N279K mutation which strengthens a splicing enhancer within E10. Conversely, increase or decrease in E10 content was achieved by trans-splicing from a target carrying the Δ280K mutation, which weakens the same splicing enhancer. Thus E10 inclusion can be modulated by trans-splicing irrespective of the strength of the cis-splicing elements affected by FTDP-17 mutations. In conclusion, RNA trans-splicing could provide the basis of therapeutic strategies for impaired alternative splicing caused by pathogenic mutations in cis-acting splicing elements.

Original languageEnglish (US)
Pages (from-to)3266-3273
Number of pages8
JournalHuman Molecular Genetics
Volume18
Issue number17
DOIs
StatePublished - 2009
Externally publishedYes

Fingerprint

Trans-Splicing
Spliceosomes
Frontotemporal Dementia
Exons
Mutation
Alternative Splicing
RNA
tau Proteins
Messenger RNA
RNA Splice Sites
Chromosomes, Human, Pair 17
COS Cells
RNA Precursors
Neurodegenerative Diseases
Introns
Transfection

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)
  • Molecular Biology

Cite this

Correction of tau mis-splicing caused by FTDP-17 MAPT mutations by spliceosome-mediated RNA trans-splicing. / Rodriguez-Martin, Teresa; Anthony, Karen; Garcia-Blanco, Mariano; Mansfield, S. Gary; Anderton, Brian H.; Gallo, Jean Marc.

In: Human Molecular Genetics, Vol. 18, No. 17, 2009, p. 3266-3273.

Research output: Contribution to journalArticle

Rodriguez-Martin, Teresa ; Anthony, Karen ; Garcia-Blanco, Mariano ; Mansfield, S. Gary ; Anderton, Brian H. ; Gallo, Jean Marc. / Correction of tau mis-splicing caused by FTDP-17 MAPT mutations by spliceosome-mediated RNA trans-splicing. In: Human Molecular Genetics. 2009 ; Vol. 18, No. 17. pp. 3266-3273.
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