title: Assessment of the feasibility of exon 45-55 multiexon skipping for Duchenne muscular dystrophy.
abstract: BACKGROUND
  The specific skipping of an exon, induced by antisense oligonucleotides (AON) during splicing, has shown to be a promising therapeutic approach for Duchenne muscular dystrophy (DMD) patients. As different mutations require skipping of different exons, this approach is mutation dependent. The skipping of an entire stretch of exons (e.g. exons 45 to 55) has recently been suggested as an approach applicable to larger groups of patients. However, this multiexon skipping approach is technically challenging. The levels of intended multiexon skips are typically low and highly variable, and may be dependent on the order of intron removal. We hypothesized that the splicing order might favor the induction of multiexon 45-55 skipping.
  
  
  METHODS
  We here tested the feasibility of inducing multiexon 45-55 in control and patient muscle cell cultures using various AON cocktails.
  
  
  RESULTS
  In all experiments, the exon 45-55 skip frequencies were minimal and comparable to those observed in untreated cells.
  
  
  CONCLUSIONS
  We conclude that current state of the art does not sufficiently support clinical development of multiexon skipping for DMD.
authors.affiliation: DMD Genetic Therapy Group, Department of Human Genetics, Leiden University, Medical Center, Postzone S4-P, PO Box 9600, 2300RC, Leiden, The Netherlands. l.van_vliet@lumc.nl
authors.forename: Laura
authors.initials: L
authors.lastname: van Vliet
authors.forename: Christa L
authors.initials: CL
authors.lastname: de Winter
authors.forename: Judith C T
authors.initials: JC
authors.lastname: van Deutekom
authors.forename: Gert-Jan B
authors.initials: GJ
authors.lastname: van Ommen
authors.forename: Annemieke
authors.initials: A
authors.lastname: Aartsma-Rus
mesh_terms: D015153:Blotting, Western; D002454:Cell Differentiation; D002478:Cells, Cultured; D005091:Exons; D015316:Genetic Therapy; D006801:Humans; D018485:Muscle Fibers, Skeletal / Q000473:pathology; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D032448:Myoblasts, Skeletal / Q000473:pathology; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D016366:Open Reading Frames; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017384:Sequence Deletion
chemical_list: D016376:Oligonucleotides, Antisense