DMPK promoter silencing by CRISPRi as a new therapeutic strategy in myotonic dystrophy type 1

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Porquet, Florent | Ormenese, S | Giroulle, F | Massotte, L | Freeman, S | Revnic, A | Gazon, H | Furling, Denis | Di Valentin, E | Gillet, N | Klein, Arnaud, F | Seutin, Vincent | Willems, Luc

Edité par CCSD -

International audience. "a. IntroductionMyotonic dystrophy type 1 (DM1) is a life threatening disease and causes severe physical and mental disabilities. Unfortunately, there are currently only symptomatic treatments. Therefore, our team aims at elaborating a new curative approach which consists in the DMPK promoter silencing by the CRISPRi system.b. MethodsThe DMPK promoter inhibition capacity of CRISPRi was tested in immortalized myoblasts from DM1 patients. For this purpose, lentiviral particles were produced using CRISPRi plasmids with their own sgRNAs. Next, these myoblasts were transduced and selected with blasticidin. Then, total DMPK mRNA was titrated by RT-qPCR and nuclear DMPK RNA foci were determinated by FISH.c. ResultsSome sgRNAs lead to near 70% inhibition of DMPK transcription as well as foci average particules in DM1 transduced myoblasts.d. ConclusionsThe CRISPRi system is able to efficiently prevent the DMPK mRNA production and foci formation in myoblast."

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