Validation and Detection of Exon Skipping Boosters in DMD Patient Cell Models and mdx Mouse

• DOI: 10.1007/978-1-4939-8651-4_19
• Publication Date: 2018-08-31T12:51:17Z
• Authors: Florian Barthelemy; Dereck Wang; Stanley F. Nelson; M. Carrie Miceli
• Keywords: Duchenne/ Becker Muscular Dystrophy; Muscular dystrophies; Intellectual and Developmental Disabilities (IDD); RNA Splicing; Muscle Fibers, Skeletal; Muscle Fibers; Dantrolene; Dystrophin; Mice; Rare Diseases; Genetics; 2.1 Biological and endogenous factors; Animals; Humans; Muscular Dystrophy; Aetiology; MyoD Protein; Pediatric; Muscle Cells; Animal; Reverse Transcriptase Polymerase Chain Reaction; Prevention; Inbred mdx; Skeletal; Exons; Genetic Therapy; Fibroblasts; Duchenne; Cellular Reprogramming; Immunohistochemistry; Brain Disorders; 3. Good health; Muscular Dystrophy, Duchenne; Disease Models, Animal; Therapies; Disease Models; Mutation; Mice, Inbred mdx; Biochemistry and Cell Biology; Other Chemical Sciences; Exon skipping; Developmental Biology

Duchenne muscular dystrophy (DMD) is caused by mutations in the DMD gene. Most deletions, duplications, or indels lead to shift of mRNA reading frame, which prevent the production of dystrophin protein. DMD is the leading fatal genetic disorder in childhood. One therapeutic strategy aims to skip one or more exons to restore reading frame to enable the production of internally truncated proteins with partial functionality. However, to date the efficiency of this strategy is suboptimal. Here we present methods for assessing exon skipping using AON alone or in combination with skip booster in the context of human DMD patient fibroblast derived myotubes and in the mdx mouse model of DMD.