Details

Autor(en) / Beteiligte

• Cardinali, Beatrice
• Provenzano, Claudia
• Izzo, Mariapaola
• Voellenkle, Christine
• Battistini, Jonathan
• Strimpakos, Georgios
• Golini, Elisabetta
• Mandillo, Silvia
• Scavizzi, Ferdinando
• Raspa, Marcello
• Perfetti, Alessandra
• Baci, Denisa
• Lazarevic, Dejan
• Garcia-Manteiga, Jose Manuel
• Gourdon, Geneviève
• Martelli, Fabio
• Falcone, Germana

Titel

Time-controlled and muscle-specific CRISPR/Cas9-mediated deletion of CTG-repeat expansion in the DMPK gene

Ist Teil von

• Molecular therapy. Nucleic acids, 2022-03, Vol.27, p.184-199

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2022

Link zum Volltext

Quelle

EZB Electronic Journals Library

Beschreibungen/Notizen

• CRISPR/Cas9-mediated therapeutic gene editing is a promising technology for durable treatment of incurable monogenic diseases such as myotonic dystrophies. Gene-editing approaches have been recently applied to in vitro and in vivo models of myotonic dystrophy type 1 (DM1) to delete the pathogenic CTG-repeat expansion located in the 3′ untranslated region of the DMPK gene. In DM1-patient-derived cells removal of the expanded repeats induced beneficial effects on major hallmarks of the disease with reduction in DMPK transcript-containing ribonuclear foci and reversal of aberrant splicing patterns. Here, we set out to excise the triplet expansion in a time-restricted and cell-specific fashion to minimize the potential occurrence of unintended events in off-target genomic loci and select for the target cell type. To this aim, we employed either a ubiquitous promoter-driven or a muscle-specific promoter-driven Cas9 nuclease and tetracycline repressor-based guide RNAs. A dual-vector approach was used to deliver the CRISPR/Cas9 components into DM1 patient-derived cells and in skeletal muscle of a DM1 mouse model. In this way, we obtained efficient and inducible gene editing both in proliferating cells and differentiated post-mitotic myocytes in vitro as well as in skeletal muscle tissue in vivo. [Display omitted] This paper describes a gene-editing approach designed to remove the pathologic CTG expansion mutation that causes myotonic dystrophy type 1. By using muscle-specific and drug-inducible expression of the CRISPR/Cas9 complex, effective deletion of the CTG expansion was obtained in myogenic cells and in mouse skeletal muscle.