Prevention of Muscular Dystrophy in Mice by Gene Editing

dc.contributor.advisorWang, Zhigaoen
dc.contributor.committeeMemberOlson, Eric N.en
dc.contributor.committeeMemberCobb, Melanie H.en
dc.contributor.committeeMemberChen, Zhijian J.en
dc.creatorLong, Chengzuen
dc.date.accessioned2017-01-03T21:46:45Z
dc.date.available2017-01-03T21:46:45Z
dc.date.created2014-12
dc.date.issued2014-11-20
dc.date.submittedDecember 2014
dc.date.updated2017-01-03T21:40:24Z
dc.description.abstractDuchenne muscular dystrophy (DMD) is an inherited X-linked disease caused by mutations in the gene encoding dystrophin, a protein required for muscle fiber integrity. DMD is characterized by progressive muscle weakness and a shortened lifespan, often along with breathing and heart complications. There is no effective treatment. RNA-guided nucleases-mediated genome editing, based on Type II CRISPR/Cas systems, offers a new approach to alter the genome. It can precisely remove a mutation in DNA, allowing the DNA repair mechanisms to replace it with a normal copy of the gene. The benefit of this over other gene therapy techniques is that it can permanently correct the 'defect' in a gene rather than just transiently adding a 'functional' one. We used CRISPR/Cas9-mediated genome editing to correct the dystrophin gene (Dmd) mutation in the germline of mdx mice, a model for DMD, and then monitored skeletal muscle and heart structure and function. Genome editing produced genetically mosaic animals containing 2 to 100% correction of the Dmd gene. Histological analysis of skeletal muscle and heart from these corrected mice showed absence of the dystrophic muscle phenotype and restoration of dystrophin expression. In addition, the degree of muscle phenotypic rescue in mosaic mice exceeded the efficiency of gene correction, likely reflecting an advantage of the corrected stem cells and their contribution to regenerating muscle. Our experiments provide proof-of-concept that CRISPR/Cas9-mediated genomic editing can correct a causative germline mutation causing muscular dystrophy in a mouse model and prevent development of several characteristic features of the disease. With rapid technological advances of gene delivery systems and improvements to the CRISPR/Cas9 editing system, this strategy may allow correction of disease-causing mutations in the muscle tissue or iPSCs (induced pluripotent stem cells) from patients with genetic diseases.en
dc.format.mimetypeapplication/pdfen
dc.identifier.oclc967597562
dc.identifier.urihttps://hdl.handle.net/2152.5/3953
dc.language.isoenen
dc.subjectCRISPR-Cas Systemsen
dc.subjectDystrophinen
dc.subjectGene Targetingen
dc.subjectMuscular Dystrophy, Duchenneen
dc.titlePrevention of Muscular Dystrophy in Mice by Gene Editingen
dc.typeThesisen
dc.type.materialtexten
thesis.degree.departmentGraduate School of Biomedical Sciencesen
thesis.degree.disciplineGenetics and Developmenten
thesis.degree.grantorUT Southwestern Medical Centeren
thesis.degree.levelDoctoralen
thesis.degree.nameDoctor of Philosophyen

Files

Original bundle

Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
LONG-DISSERTATION-2014.pdf
Size:
4.36 MB
Format:
Adobe Portable Document Format

License bundle

Now showing 1 - 1 of 1
No Thumbnail Available
Name:
LICENSE.txt
Size:
1.84 KB
Format:
Plain Text
Description: