U7 snRNAs induce correction of mutated dystrophin pre-mRNA by exon skipping.
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.
Brun
C
C
Division of Neuropathology, Institute of Pathology, University of Bern, Murtenstrasse 31, 3010 Bern, Switzerland.
Suter
D
D
Pauli
C
C
Dunant
P
P
Lochmüller
H
H
Burgunder
J M
JM
Schümperli
D
D
Weis
J
J
D000818:Animals; D016189:Dystrophin / Q000378:metabolism*; D005091:Exons; D015316:Genetic Therapy; D051379:Mice; D018101:Mice, Inbred mdx; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000628:therapy; D009154:Mutation; D012322:RNA Precursors / Q000378:metabolism*; D012326:RNA Splicing / Q000502:physiology*; D016372:RNA, Antisense / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000378:metabolism*
D016189:Dystrophin; D012322:RNA Precursors; D016372:RNA, Antisense; D012342:RNA, Small Nuclear; C405487:U7 small nuclear RNA