Specific DMPK-promoter targeting by CRISPRi reverses myotonic dystrophy type 1-associated defects in patient muscle cells

Myotonic dystrophy type 1 (DM1) is a neuromuscular disease that originates from an expansion of CTG microsatellites in the 3′ untranslated region of the DMPK gene, thus leading to the expression of transcripts containing expanded CUG repeats (CUGexp). The pathophysiology is explained by a toxic RNA...

Full description

Saved in:

Bibliographic Details
Published in:	Molecular therapy. Nucleic acids Vol. 32; pp. 857 - 871
Main Authors:	Porquet, Florent, Weidong, Lin, Jehasse, Kévin, Gazon, Hélène, Kondili, Maria, Blacher, Silvia, Massotte, Laurent, Di Valentin, Emmannuel, Furling, Denis, Gillet, Nicolas Albert, Klein, Arnaud François, Seutin, Vincent, Willems, Luc
Format:	Journal Article Web Resource
Language:	English
Published:	United States Elsevier Inc 13-06-2023 Elsevier Cell Press American Society of Gene & Cell Therapy
Subjects:	Biochemistry, Molecular Biology CRISPRi Drug Discovery gene silencing gene therapy Human health sciences Life Sciences Molecular biology Molecular Medicine MT: RNA/DNA Editing myotonic dystrophy type 1 Neurologie Neurology neuromuscular disease Original Sciences de la santé humaine gene silencing MT: RNA/DNA Editing gene therapy neuromuscular disease CRISPRi myotonic dystrophy type 1 Modèles cellulaires Cellular model Myotonic Dystrophy Dystrophie Mytotonique de type 1 Thérapie génique
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Myotonic dystrophy type 1 (DM1) is a neuromuscular disease that originates from an expansion of CTG microsatellites in the 3′ untranslated region of the DMPK gene, thus leading to the expression of transcripts containing expanded CUG repeats (CUGexp). The pathophysiology is explained by a toxic RNA gain of function where CUGexp RNAs form nuclear aggregates that sequester and alter the function of MBNL splicing factors, triggering splicing misregulation linked to the DM1 symptoms. There is currently no cure for DM1, and most therapeutic strategies aim at eliminating CUGexp-DMPK transcripts. Here, we investigate a DMPK-promoter silencing strategy using CRISPR interference as a new alternative approach. Different sgRNAs targeting the DMPK promoter are evaluated in DM1 patient muscle cells. The most effective guides allowed us to reduce the level of DMPK transcripts and CUGexp-RNA aggregates up to 80%. The CUGexp-DMPK repression corrects the overall transcriptome, including spliceopathy, and reverses a physiological parameter in DM1 muscle cells. Its action is specific and restricted to the DMPK gene, as confirmed by genome-wide expression analysis. Altogether, our findings highlight DMPK-promoter silencing by CRISPRi as a promising therapeutic approach for DM1. [Display omitted] Porquet and colleagues develop an alternative therapeutic approach for DM1. They show that targeting of the DMPK promoter by CRISPR interference reduces the level of the toxic CUGexp-DMPK transcripts, thus improving the DM1 hallmarks, including foci formation and splicing defects, along with high transcriptomic specificity in human muscle cells.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 scopus-id:2-s2.0-85159927755 These authors contributed equally
ISSN:	2162-2531 2162-2531
DOI:	10.1016/j.omtn.2023.05.007