Myospreader improves gene editing in skeletal muscle by myonuclear propagation.
Proc Natl Acad Sci U S A
; 121(19): e2321438121, 2024 May 07.
Article
in En
| MEDLINE
| ID: mdl-38687782
ABSTRACT
Successful CRISPR/Cas9-based gene editing in skeletal muscle is dependent on efficient propagation of Cas9 to all myonuclei in the myofiber. However, nuclear-targeted gene therapy cargos are strongly restricted to their myonuclear domain of origin. By screening nuclear localization signals and nuclear export signals, we identify "Myospreader," a combination of short peptide sequences that promotes myonuclear propagation. Appending Myospreader to Cas9 enhances protein stability and myonuclear propagation in myoblasts and myofibers. AAV-delivered Myospreader dCas9 better inhibits transcription of toxic RNA in a myotonic dystrophy mouse model. Furthermore, Myospreader Cas9 achieves higher rates of gene editing in CRISPR reporter and Duchenne muscular dystrophy mouse models. Myospreader reveals design principles relevant to all nuclear-targeted gene therapies and highlights the importance of the spatial dimension in therapeutic development.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
Genetic Therapy
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Cell Nucleus
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Muscle, Skeletal
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Muscular Dystrophy, Duchenne
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CRISPR-Cas Systems
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Gene Editing
Limits:
Animals
/
Humans
Language:
En
Year:
2024
Type:
Article