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

Florian Barthelemy; Dereck Wang; Stanley F Nelson; M Carrie Miceli

doi:10.1007/978-1-4939-8651-4_19

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

Methods Mol Biol. 2018:1828:309-326. doi: 10.1007/978-1-4939-8651-4_19.

Authors

Florian Barthelemy^{1

2}, Dereck Wang^{1

2}, Stanley F Nelson^{1

3

4}, M Carrie Miceli^{5

6

7}

Affiliations

¹ Center for Duchenne Muscular Dystrophy, University of California, Los Angeles, Los Angeles, CA, USA.
² Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, USA.
³ Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
⁴ Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
⁵ Center for Duchenne Muscular Dystrophy, University of California, Los Angeles, Los Angeles, CA, USA. cmiceli@ucla.edu.
⁶ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, USA. cmiceli@ucla.edu.
⁷ Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine and College of Letters and Sciences, University of California, Los Angeles, Los Angeles, CA, USA. cmiceli@ucla.edu.

PMID: 30171550
DOI: 10.1007/978-1-4939-8651-4_19

Abstract

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.

Keywords: Dantrolene; Dystrophin; Exon skipping; Muscular dystrophies; Therapies.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cellular Reprogramming / genetics
Disease Models, Animal
Dystrophin / genetics*
Exons*
Fibroblasts / metabolism
Genetic Therapy
Humans
Immunohistochemistry
Mice
Mice, Inbred mdx
Muscle Cells / metabolism
Muscle Fibers, Skeletal / metabolism
Muscular Dystrophy, Duchenne / genetics*
Muscular Dystrophy, Duchenne / therapy
Mutation
MyoD Protein / genetics
MyoD Protein / metabolism
RNA Splicing*
Reverse Transcriptase Polymerase Chain Reaction

Substances

Dystrophin
MyoD Protein

Abstract

Publication types

MeSH terms

Substances

Grants and funding