Reprogramming of tau alternative splicing by spliceosome-mediated RNA trans-splicing: Implications for tauopathies

Teresa Rodriguez-Martin, Mariano A. Garcia-Blanco, S. Gary Mansfield, Andrew C. Grower, Michael Hutton, Qingming Yu, Jianhua Zhou, Brian H. Anderten, Jean Marc Gallo

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17) is caused by mutations in the gene encoding the microtubule-associated protein, tau. Some FTDP-17 mutations affect exon 10 splicing. To correct aberrant exon 10 splicing while retaining endogenous transcriptional control, we evaluated the feasibility of using spliceosome-mediated RNA trans-splicing (SMaRT) to reprogram tau mRNA. We designed a pre-trans-splicing molecule containing human tau exons 10 to 13 and a binding domain complementary to the 3′ end of tau intron 9. A minigene comprising tau exons 9, 10, and 11 and minimal flanking intronic sequences was used as a target. RT-PCR analysis of SH-SYSY cells or COS cells cotransfected with a minigene and a pre-trans-splicing molecule using primers to opposite sides of the predicted splice junction generated products containing exons 9 to 13. Sequencing of the chimeric products showed that an exact exon 9-exon 10 junction had been created, thus demonstrating that tau RNA can be reprogrammed by trans-splicing. Furthermore, by using the same paradigm with a minigene containing full-length intronic sequences, we show that cis-splicing exclusion of exon 10 can be by-passed by trans-splicing and that conversion of exon 10- tau RNA into exon 10+ tau RNA could be achieved with ≈34% efficiency. Our results demonstrate that an alternatively spliced exon can be replaced by trans-splicing and open the way to novel therapeutic applications of SMaRT for tauopathies and other disorders linked to aberrant alternative splicing.

Original languageEnglish
Pages (from-to)15659-15664
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume102
Issue number43
DOIs
StatePublished - Oct 25 2005
Externally publishedYes

Fingerprint

Trans-Splicing
Tauopathies
Spliceosomes
Alternative Splicing
Exons
Frontotemporal Dementia
RNA
Chromosomes, Human, Pair 17
Mutation
Microtubule-Associated Proteins
COS Cells
Introns

Keywords

  • Cytoskeleton
  • Dementia
  • Neurodegeneration
  • RNA therapy

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Reprogramming of tau alternative splicing by spliceosome-mediated RNA trans-splicing : Implications for tauopathies. / Rodriguez-Martin, Teresa; Garcia-Blanco, Mariano A.; Mansfield, S. Gary; Grower, Andrew C.; Hutton, Michael; Yu, Qingming; Zhou, Jianhua; Anderten, Brian H.; Gallo, Jean Marc.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 102, No. 43, 25.10.2005, p. 15659-15664.

Research output: Contribution to journalArticle

Rodriguez-Martin, Teresa ; Garcia-Blanco, Mariano A. ; Mansfield, S. Gary ; Grower, Andrew C. ; Hutton, Michael ; Yu, Qingming ; Zhou, Jianhua ; Anderten, Brian H. ; Gallo, Jean Marc. / Reprogramming of tau alternative splicing by spliceosome-mediated RNA trans-splicing : Implications for tauopathies. In: Proceedings of the National Academy of Sciences of the United States of America. 2005 ; Vol. 102, No. 43. pp. 15659-15664.
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