CRISPR/dCas9-mediated transcriptional inhibition ameliorates the epigenetic dysregulation at D4Z4 and represses DUX4-fl in FSH muscular dystrophy

UMMS Affiliation

Department of Cell and Developmental Biology; Department of Neurology

Publication Date


Document Type



Cell Biology | Developmental Biology | Molecular Biology | Molecular Genetics | Musculoskeletal Diseases | Nervous System Diseases


Facioscapulohumeral muscular dystrophy (FSHD) is one of the most prevalent myopathies, affecting males and females of all ages. Both forms of the disease are linked by epigenetic de-repression of the D4Z4 macrosatellite repeat array at chromosome 4q35, leading to aberrant expression of D4Z4-encoded RNAs in skeletal muscle. Production of full-length DUX4 (DUX4-fl) mRNA from the de-repressed D4Z4 array results in misexpression of DUX4-FL protein and its transcriptional targets, and apoptosis, ultimately leading to accumulated muscle pathology. Returning the chromatin at the FSHD locus to its non-pathogenic, epigenetically repressed state would simultaneously affect all D4Z4 RNAs, inhibiting downstream pathogenic pathways, and is thus an attractive therapeutic strategy. Advances in CRISPR/Cas9-based genome editing make it possible to target epigenetic modifiers to an endogenous disease locus, although reports to date have focused on more typical genomic regions. Here we demonstrate that a CRISPR/dCas9 transcriptional inhibitor can be specifically targeted to the highly repetitive FSHD macrosatellite array and alter the chromatin to repress expression of DUX4-fl in primary FSHD myocytes. These results implicate the promoter and exon 1 of DUX4 as potential therapeutic targets and demonstrate the utility of CRISPR technology for correction of the epigenetic dysregulation in FSHD.

DOI of Published Version



Himeda CL, Jones TI, Jones PL. CRISPR/dCas9-mediated Transcriptional Inhibition Ameliorates the Epigenetic Dysregulation at D4Z4 and Represses DUX4-fl in FSH Muscular Dystrophy. Mol Ther. 2016 Mar;24(3):527-35. doi:10.1038/mt.2015.200. Epub 2015 Nov 3. PubMed PMID: 26527377; PubMed Central PMCID: PMC4786914. Link to article on publisher's website

Journal/Book/Conference Title

Molecular therapy : the journal of the American Society of Gene Therapy


Acknowledgments: This work was financially supported by the National Institute of Arthritis, Musculoskeletal, and Skin Diseases grant #1R01AR062587 and the Association Française contre les Myopathies grant #AFM15700. The authors thank Kathryn R. Wagner and the UMMS Wellstone Center for providing cells, and the Chris Carrino Foundation for FSHD for their support of our FSHD research projects. The authors declare no conflicts of interest.

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