Human Molecular Genetics Advance Access published online on June 4, 2009
Human Molecular Genetics, doi:10.1093/hmg/ddp264
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Correction of tau mis-splicing caused by FTDP-17 MAPT mutations by spliceosome-mediated RNA trans-splicing
1 MRC Centre for Neurodegeneration Research, King's College London, Institute of Psychiatry, Department of Clinical Neuroscience, De Crespigny Park, London SE5 8AF, United Kingdom 2 Center for RNA Biology, Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA 3 VIRxSYS Inc. 200 Perry Parkway, Gaithersburg, MD 20877, USA
* Corresponding author: Jean-Marc Gallo MRC Centre for Neurodegeneration Research, King's College London, Institute of Psychiatry, Department of Clinical Neuroscience, Box PO37, De Crespigny Park, London, SE5 8AF, United Kingdom. Tel. No. +44 207 848 0404; Fax No. +44 207 708 0017, E-mail: jean-marc.gallo{at}iop.kcl.ac.uk
Received May 5, 2009; Revised May 5, 2009; Accepted June 1, 2009
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 were 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 strengthening a splicing enhancer within E10. Conversely, increase or decrease in E10 content was achieved by trans-splicing from a target carrying the 
280K mutation, that 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.