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Fast, multiplexable and efficient somatic gene deletions in adult mouse skeletal muscle fibers using AAV-CRISPR/Cas9

Molecular screens comparing different disease states to identify candidate genes rely on the availability of fast, reliable and multiplexable systems to interrogate genes of interest. CRISPR/Cas9-based reverse genetics is a promising method to eventually achieve this. However, such methods are sorel...

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Bibliographic Details
Main Authors: Thürkauf, Marco (Author) , Lin, Shuo (Author) , Oliveri, Filippo (Author) , Grimm, Dirk (Author) , Platt, Randall J. (Author) , Rüegg, Markus A. (Author)
Format: Article (Journal)
Language:English
Published: 30 September 2023
In: Nature Communications
Year: 2023, Volume: 14, Pages: 1-16
ISSN:2041-1723
DOI:10.1038/s41467-023-41769-7
Online Access:Verlag, kostenfrei, Volltext: https://doi.org/10.1038/s41467-023-41769-7
Verlag, kostenfrei, Volltext: https://www.nature.com/articles/s41467-023-41769-7
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Author Notes:Marco Thürkauf, Shuo Lin, Filippo Oliveri, Dirk Grimm, Randall J. Platt & Markus A. Rüegg
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Summary:Molecular screens comparing different disease states to identify candidate genes rely on the availability of fast, reliable and multiplexable systems to interrogate genes of interest. CRISPR/Cas9-based reverse genetics is a promising method to eventually achieve this. However, such methods are sorely lacking for multi-nucleated muscle fibers, since highly efficient nuclei editing is a requisite to robustly inactive candidate genes. Here, we couple Cre-mediated skeletal muscle fiber-specific Cas9 expression with myotropic adeno-associated virus-mediated sgRNA delivery to establish a system for highly effective somatic gene deletions in mice. Using well-characterized genes, we show that local or systemic inactivation of these genes copy the phenotype of traditional gene-knockout mouse models. Thus, this proof-of-principle study establishes a method to unravel the function of individual genes or entire signaling pathways in adult skeletal muscle fibers without the cumbersome requirement of generating knockout mice.
Item Description:Gesehen am 11.03.2024
Physical Description:Online Resource
ISSN:2041-1723
DOI:10.1038/s41467-023-41769-7

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