Cellular and Molecular Life Sciences CMLS

, Volume 60, Issue 3, pp 557–566

U7 snRNAs induce correction of mutated dystrophin pre-mRNA by exon skipping

  • C. Brun
  • D. Suter
  • C. Pauli
  • P. Dunant
  • H. Lochmüller
  • J.-M. Burgunder
  • D. Schümperli
  • J. Weis
Research Article

DOI: 10.1007/s000180300047

Cite this article as:
Brun, C., Suter, D., Pauli, C. et al. CMLS, Cell. Mol. Life Sci. (2003) 60: 557. doi:10.1007/s000180300047

Abstract.

Most cases of Duchenne muscular dystrophy are caused by dystrophin gene mutations that disrupt the mRNA reading frame. Artificial exclusion (skipping) of a single exon would often restore the reading frame, giving rise to a shorter, but still functional dystrophin protein. Here, we analyzed the ability of antisense U7 small nuclear (sn)RNA derivatives to alter dystrophin pre-mRNA splicing. As a proof of principle, we first targeted the splice sites flanking exon 23 of dystrophin pre-mRNA in the wild-type muscle cell line C2C12 and showed precise exon 23 skipping. The same strategy was then successfully adapted to dystrophic immortalized mdx muscle cells where exon-23-skipped dystrophin mRNA rescued dystrophin protein synthesis. Moreover, we observed a stimulation of antisense U7 snRNA expression by the murine muscle creatine kinase enhancer. These results demonstrate that alteration of dystrophin pre-mRNA splicing could correct dystrophin gene mutations by expression of specific U7 snRNA constructs.

Key words. Duchenne muscular dystrophy; dystrophin; exon skipping; gene therapy; pre-mRNA; U7 snRNA.

Copyright information

© Birkhäuser Verlag, 2003

Authors and Affiliations

  • C. Brun
    • 1
  • D. Suter
    • 1
  • C. Pauli
    • 1
  • P. Dunant
    • 2
  • H. Lochmüller
    • 2
  • J.-M. Burgunder
    • 3
  • D. Schümperli
    • 4
  • J. Weis
    • 1
  1. 1.Division of Neuropathology, Institute of Pathology, University of Bern, Murtenstrasse 31, 3010 Bern (Switzerland), Fax +41 31 632 9872, e-mail: joachim.weis@pathology.unibe.chCH
  2. 2.Gene Center and Friedrich-Baur-Institute, Ludwig-Maximilians-University, Munich (Germany)DE
  3. 3.Department of Neurology, Inselspital University Hospital, Bern (Switzerland)CH
  4. 4.Institute of Cell Biology, University of Bern, Bern (Switzerland)CH