Creation of a Novel Humanized Dystrophic Mouse Model of Duchenne Muscular Dystrophy and Application of a CRISPR/Cas9 Gene Editing Therapy

Courtney S Young; Ekaterina Mokhonova; Marbella Quinonez; April D Pyle; Melissa J Spencer

doi:10.3233/JND-170218

Creation of a Novel Humanized Dystrophic Mouse Model of Duchenne Muscular Dystrophy and Application of a CRISPR/Cas9 Gene Editing Therapy

J Neuromuscul Dis. 2017;4(2):139-145. doi: 10.3233/JND-170218.

Authors

Courtney S Young^{1

2

3}, Ekaterina Mokhonova^{2

4}, Marbella Quinonez^{2

5}, April D Pyle^{2

3

6}, Melissa J Spencer^{1

2

3

4}

Affiliations

¹ Molecular Biology Interdepartmental Program, University of California, Los Angeles, CA, USA.
² Center for Duchenne Muscular Dystrophy at UCLA, University of California, Los Angeles, CA, USA.
³ Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, LosAngeles, CA, USA.
⁴ Department of Neurology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
⁵ Department of Physiology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.
⁶ Department of Microbiology, Immunology and Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.

Abstract

Duchenne muscular dystrophy is caused by mutations in DMD which disrupt the reading frame. Therapeutic strategies that restore DMD's reading frame, such as exon skipping and CRISPR/Cas9, need to be tested in the context of the human DMD sequence in vivo. We have developed a novel dystrophic mouse model by using CRISPR/Cas9 to delete exon 45 in the human DMD gene in hDMD mice, which places DMD out-of-frame. We have utilized this model to demonstrate that our clinically-relevant CRISPR/Cas9 platform, which targets deletion of human DMD exons 45-55, can be directly applied in vivo to restore dystrophin.

Keywords: CRISPR; Duchenne muscular dystrophy; animal models; gene editing; mice.

MeSH terms

Animals
CRISPR-Cas Systems
Disease Models, Animal*
Dystrophin / genetics*
Dystrophin / metabolism
Exons
Gene Editing / methods
Genetic Therapy* / methods
HEK293 Cells
Humans
Mice, Transgenic
Muscle, Skeletal / metabolism
Muscle, Skeletal / pathology
Muscular Dystrophy, Duchenne / genetics
Muscular Dystrophy, Duchenne / metabolism
Muscular Dystrophy, Duchenne / pathology
Muscular Dystrophy, Duchenne / therapy*

Substances

DMD protein, human
Dystrophin

Abstract

MeSH terms

Substances

Grants and funding