Switch-off the trouble:DMPK promoter targeting by CRISPRi as an original specific therapy in DM1 - Repeat Expansions & Myotonic Dystrophy (REDs) Accéder directement au contenu
Poster De Conférence Année : 2022

Switch-off the trouble:DMPK promoter targeting by CRISPRi as an original specific therapy in DM1

Résumé

"Introduction: Myotonic dystrophy type 1 (DM1) originates from an amplification of CTG microsatellites in the DMPK gene. The pathology is primarily explained by a toxic gain of function where the expanded-CUG DMPK transcripts induce mainly the loss of function of the MBNL proteins, triggering a wide spliceopathy. Several therapies have been tested to neutralize the toxic DMPK transcripts or their consequences. Here, we investigated a new therapeutic strategy consisting of the silencing of the DMPK promoter by a CRISPRi system in patient-derived myotubes. Methods: Our DMPK repressing strategy by CRISPRi was tested in immortalized myoblasts from a DM1 patient or a healthy donor. Stable cell lines expressing a deactivated Cas9 conjugated to an inhibitory KRAB domain in addition to their own sgRNAs were produced by lentiviral transduction. The efficacy and specificity of our therapeutic strategy were then assessed in differentiated myotubes. Results: Our DMPK promoter inhibition strategy is highly efficient to reduce toxic DMPK transcript quantities up to 80%. This level of inhibition allows to correct the DM1 hallmark defects by reducing the presence of foci, improving the spliceopathy and normalizing an electrophysiological parameter in DM1 myotubes. Furthermore, this approach displays unprecedented high specificity as evidenced by a complete lack of off-target effects on the transcriptome from unaffected myotubes. Conclusions: We conclude that DMPK promoter inhibition is a promising strategy to be developed for DM1 treatment."
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Dates et versions

hal-04022715 , version 1 (10-03-2023)

Identifiants

  • HAL Id : hal-04022715 , version 1

Citer

Florent Porquet, L Weidong, K Jehasse, H Gazon, Maria Kondili, et al.. Switch-off the trouble:DMPK promoter targeting by CRISPRi as an original specific therapy in DM1. 7th international congress of Myology, Sep 2022, Nice, France. ⟨hal-04022715⟩
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