title: Efficient and fast functional screening of microdystrophin constructs in vivo and in vitro for therapy of duchenne muscular dystrophy.
abstract: Duchenne muscular dystrophy (DMD) is an X-linked, lethal genetic disorder affecting the skeletal muscle compartment, and is caused by mutation(s) in the dystrophin gene. Gene delivery of microdystrophin constructs using adeno-associated virus (AAV) and antisense-mediated exon skipping restoring the genetic reading frame are two of the most promising therapeutic strategies for DMD. Both approaches use microdystrophin proteins either directly as a desired construct for gene delivery, using the capacity-limited AAV vectors, or as the therapeutic outcome of gene splicing. Although functionality of the resulting artificial dystrophin proteins can be predicted in silico, experimental evidence usually obtained in transgenic mice is required before human trials. However, the enormous number of potential constructs makes screening assays for dystrophin protein function in vitro and in vivo highly desirable. Here we present data showing that functionality of microdystrophins can be assessed using relatively simple and fast techniques.
authors.affiliation: Institute of Human Genetics, University of Newcastle, Newcastle upon Tyne NE13BZ, United Kingdom.
authors.forename: Louise H
authors.initials: LH
authors.lastname: Jørgensen
authors.forename: Nancy
authors.initials: N
authors.lastname: Larochelle
authors.forename: Kristian
authors.initials: K
authors.lastname: Orlopp
authors.forename: Patrick
authors.initials: P
authors.lastname: Dunant
authors.forename: Roy W R
authors.initials: RW
authors.lastname: Dudley
authors.forename: Rolf
authors.initials: R
authors.lastname: Stucka
authors.forename: Christian
authors.initials: C
authors.lastname: Thirion
authors.forename: Maggie C
authors.initials: MC
authors.lastname: Walter
authors.forename: Steven H
authors.initials: SH
authors.lastname: Laval
authors.forename: Hanns
authors.initials: H
authors.lastname: Lochmüller
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D016189:Dystrophin / Q000737:chemistry / Q000235:genetics* / Q000627:therapeutic use*; D015316:Genetic Therapy; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D018101:Mice, Inbred mdx; D009119:Muscle Contraction; D018482:Muscle, Skeletal / Q000473:pathology; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000503:physiopathology / Q000628:therapy*; D032446:Myoblasts / Q000378:metabolism / Q000473:pathology; D049031:Sarcoglycans / Q000378:metabolism; D014162:Transfection
chemical_list: D016189:Dystrophin; D049031:Sarcoglycans