Efficient CRISPR/Cas9-mediated editing of trinucleotide repeat expansion in myotonic dystrophy patient-derived iPS and myogenic cells

Sumitava Dastidar, Simon Ardui, Kshitiz Singh, Debanjana Majumdar, Nisha Nair Shivsankaran Nair, Yanfang Fu, Deepak Reyon, Ermira Samara, Mattia FM Gerbil, Arnaud F. Klein, Wito De Schrijver, Jaitip Tipanee, Sara Seneca, Warut Tulalamba, Hui Wang, Yoke Chin Chai, Pieter In 'T Veld, Denis Furling, Francesco Saverio Tedesco, Joris R. VermeeschJ. Keith Joung, Marinee Chuah, Thierry VandenDriessche

Research output: Contribution to journalArticlepeer-review

62 Citations (Scopus)


CRISPR/Cas9 is an attractive platform to potentially correct dominant genetic diseases by gene editing with unprecedented precision. In the current proof-of-principle study, we explored the use of CRISPR/Cas9 for gene-editing in myotonic dystrophy type-1 (DM1), an autosomal-dominant muscle disorder, by excising the CTG-repeat expansion in the 3'-untranslated-region (UTR) of the human myotonic dystrophy protein kinase (DMPK) gene in DM1 patient-specific induced pluripotent stem cells (DM1-iPSC), DM1-iPSC-derived myogenic cells and DM1 patient-specific myoblasts. To eliminate the pathogenic gain-of-function mutant DMPK transcript, we designed a dual guide RNA based strategy that excises the CTG-repeat expansion with high efficiency, as confirmed by Southern blot and single molecule real-time (SMRT) sequencing. Correction efficiencies up to 90% could be attained in DM1-iPSC as confirmed at the clonal level, following ribonucleoprotein (RNP) transfection of CRISPR/Cas9 components without the need for selective enrichment. Expanded CTG repeat excision resulted in the disappearance of ribonuclear foci, a quintessential cellular phenotype of DM1, in the corrected DM1-iPSC, DM1-iPSC-derived myogenic cells and DM1 myoblasts. Consequently, the normal intracellular localization of the muscleblind-like splicing regulator 1 (MBNL1) was restored, resulting in the normalization of splicing pattern of SERCA1. This study validates the use of CRISPR/Cas9 for gene editing of repeat expansions.
Original languageEnglish
Pages (from-to)8275-8298
Number of pages24
JournalNucleic Acids Research
Issue number16
Publication statusPublished - 19 Sep 2018


  • myotonic dystrophy
  • myogenic cells
  • myotonic dystrophy patient-derived iPS


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