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Live‐imaging of revertant and therapeutically restored dystrophin in the Dmd EGFP‐mdx mouse model for Duchenne muscular dystrophy
- Source :
- Neuropathology and Applied Neurobiology, Neuropathology and Applied Neurobiology, Wiley, 2020, 46 (6), pp.602-614. ⟨10.1111/nan.12639⟩, Neuropathology and Applied Neurobiology, 2020, 46 (6), pp.602-614. ⟨10.1111/nan.12639⟩
- Publication Year :
- 2020
- Publisher :
- HAL CCSD, 2020.
-
Abstract
- International audience; Background: Dmdmdx, harbouring the c.2983C>T nonsense mutation in Dmd exon 23, is a mouse model for Duchenne muscular dystrophy (DMD), frequently used to test therapies aimed at dystrophin restoration. Current translational research is methodologically hampered by the lack of a reporter mouse model, which would allow direct visualization of dystrophin expression as well as longitudinal in vivo studies. Methods: We generated a DmdEGFP-mdx reporter allele carrying in cis the mdx-23 mutation and a C-terminal EGFP-tag. This mouse model allows direct visualization of spontaneously and therapeutically restored dystrophin-EGFP fusion protein either after natural fibre reversion, or for example, after splice modulation using tricyclo-DNA to skip Dmd exon 23, or after gene editing using AAV-encoded CRISPR/Cas9 for Dmd exon 23 excision. Results: Intravital microscopy in anaesthetized mice allowed live-imaging of sarcolemmal dystrophin-EGFP fusion protein of revertant fibres as well as following therapeutic restoration. Dystrophin-EGFP-fluorescence persisted ex vivo, allowing live-imaging of revertant and therapeutically restored dystrophin in isolated fibres ex vivo. Expression of the shorter dystrophin-EGFP isoforms Dp71 in the brain, Dp260 in the retina, and Dp116 in the peripheral nerve remained unabated by the mdx-23 mutation. Conclusion: Intravital imaging of DmdEGFP-mdx muscle permits novel experimental approaches such as the study of revertant and therapeutically restored dystrophin in vivo and ex vivo.
- Subjects :
- 0301 basic medicine
musculoskeletal diseases
Duchenne muscular dystrophy
mdx mouse
congenital, hereditary, and neonatal diseases and abnormalities
tcDNA
Histology
[SDV.BIO]Life Sciences [q-bio]/Biotechnology
[SDV]Life Sciences [q-bio]
Nonsense mutation
Biology
revertant muscle fibre
Pathology and Forensic Medicine
03 medical and health sciences
Exon
0302 clinical medicine
In vivo
Physiology (medical)
medicine
CRISPR/Cas9
dystrophin-EGFP fusion protein
ComputingMilieux_MISCELLANEOUS
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology
medicine.disease
musculoskeletal system
mdx reporter mouse model
Cell biology
[SDV] Life Sciences [q-bio]
030104 developmental biology
Neurology
[SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics
biology.protein
Neurology (clinical)
Dystrophin
600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit
030217 neurology & neurosurgery
Ex vivo
Intravital microscopy
Subjects
Details
- Language :
- English
- ISSN :
- 03051846 and 13652990
- Database :
- OpenAIRE
- Journal :
- Neuropathology and Applied Neurobiology, Neuropathology and Applied Neurobiology, Wiley, 2020, 46 (6), pp.602-614. ⟨10.1111/nan.12639⟩, Neuropathology and Applied Neurobiology, 2020, 46 (6), pp.602-614. ⟨10.1111/nan.12639⟩
- Accession number :
- edsair.doi.dedup.....1ac4dc4eb39c84dc5f560636937ef89d