Gene Editing Targeting the DUX4 Polyadenylation Signal: A Therapy for FSHD?

Romains Joubert; Virginie Mariot; Marine Charpentier; Jean Paul Concordet; Julie Dumonceaux

doi:10.3390/jpm11010007

Gene Editing Targeting the DUX4 Polyadenylation Signal: A Therapy for FSHD?

J Pers Med. 2020 Dec 23;11(1):7. doi: 10.3390/jpm11010007.

Authors

Romains Joubert¹, Virginie Mariot¹, Marine Charpentier², Jean Paul Concordet², Julie Dumonceaux^{1

3}

Affiliations

¹ NIHR Biomedical Research Centre, University College London, Great Ormond Street Institute of Child Health and Great Ormond Street Hospital NHS Trust, London WC1N 1EH, UK.
² Genome Structure and Instability laboratory, CNRS UMR 7196, Inserm U1154, National Museum of Natural History, 75005 Paris, France.
³ Northern Ireland Center for Stratified Medicine, Biomedical Sciences Research Institute, Londonderry BT47 6SB, UK.

Abstract

Facioscapulohumeral dystrophy (FSHD, OMIM: 158900, 158901) is the most common dystrophy in adults and so far, there is no treatment. Different loci of the disease have been characterized and they all lead to the aberrant expression of the DUX4 protein, which impairs the function of the muscle, ultimately leading to cell death. Here, we used gene editing to try to permanently shut down DUX4 expression by targeting its poly(A) sequence. We used transcription activator-like effector nucleases (TALEN) and CRISPR-Cas9 nucleases in vitro on FSHD myoblasts. More than 150 TOPO clones were sequenced and only indels were observed in 4%. Importantly, in 2 of them, the DUX4 poly(A) signal was eliminated at the genomic level but DUX4 mRNA was still produced thanks to the use of a non-canonical upstream poly(A) signal sequence. These experiments show that targeting DUX4 PAS at the genomic level might not be an appropriate gene editing strategy for FSHD therapy.

Keywords: CRISPR-Cas9; D4Z4; DUX4; FSHD; TALEN; facioscapulohumeral dystrophy; gene editing; muscle; polyadenylation.

Abstract

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