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In vivo genome editing in mouse restores dystrophin expression in Duchenne muscular dystrophy patient muscle fibers
- Source :
- Genome Medicine, Vol 13, Iss 1, Pp 1-19 (2021), Genome Medicine
- Publication Year :
- 2021
- Publisher :
- BMC, 2021.
-
Abstract
- Background Mutations in the DMD gene encoding dystrophin—a critical structural element in muscle cells—cause Duchenne muscular dystrophy (DMD), which is the most common fatal genetic disease. Clustered regularly interspaced short palindromic repeat (CRISPR)-mediated gene editing is a promising strategy for permanently curing DMD. Methods In this study, we developed a novel strategy for reframing DMD mutations via CRISPR-mediated large-scale excision of exons 46–54. We compared this approach with other DMD rescue strategies by using DMD patient-derived primary muscle-derived stem cells (DMD-MDSCs). Furthermore, a patient-derived xenograft (PDX) DMD mouse model was established by transplanting DMD-MDSCs into immunodeficient mice. CRISPR gene editing components were intramuscularly delivered into the mouse model by adeno-associated virus vectors. Results Results demonstrated that the large-scale excision of mutant DMD exons showed high efficiency in restoring dystrophin protein expression. We also confirmed that CRISPR from Prevotella and Francisella 1(Cas12a)-mediated genome editing could correct DMD mutation with the same efficiency as CRISPR-associated protein 9 (Cas9). In addition, more than 10% human DMD muscle fibers expressed dystrophin in the PDX DMD mouse model after treated by the large-scale excision strategies. The restored dystrophin in vivo was functional as demonstrated by the expression of the dystrophin glycoprotein complex member β-dystroglycan. Conclusions We demonstrated that the clinically relevant CRISPR/Cas9 could restore dystrophin in human muscle cells in vivo in the PDX DMD mouse model. This study demonstrated an approach for the application of gene therapy to other genetic diseases.
- Subjects :
- Male
musculoskeletal diseases
Duchenne muscular dystrophy
congenital, hereditary, and neonatal diseases and abnormalities
CRISPR/ Cas12a
Genetic enhancement
Muscle Fibers, Skeletal
Gene editing
QH426-470
medicine.disease_cause
Dystrophin
Mice
Exon
Genome editing
medicine
Genetics
Animals
Humans
CRISPR
Amino Acid Sequence
Molecular Biology
CRISPR/Cas9
Genetics (clinical)
Mutation
Genome
Base Sequence
biology
Cas9
Patient-derived xenograft model
Research
Cell Differentiation
medicine.disease
Cell biology
Muscular Dystrophy, Duchenne
Disease Models, Animal
HEK293 Cells
Child, Preschool
biology.protein
Molecular Medicine
Medicine
CRISPR-Cas Systems
Transcriptome
Subjects
Details
- Language :
- English
- Volume :
- 13
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Genome Medicine
- Accession number :
- edsair.doi.dedup.....32d3202590dbf02cd90b5d2630dd66e0