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  • Targeted genome editing in vivo corrects a Dmd duplication restoring wild-type dystrophin expression.

Targeted genome editing in vivo corrects a Dmd duplication restoring wild-type dystrophin expression.

EMBO molecular medicine (2021-03-17)
Eleonora Maino, Daria Wojtal, Sonia L Evagelou, Aiman Farheen, Tatianna W Y Wong, Kyle Lindsay, Ori Scott, Samar Z Rizvi, Elzbieta Hyatt, Matthew Rok, Shagana Visuvanathan, Amanda Chiodo, Michelle Schneeweiss, Evgueni A Ivakine, Ronald D Cohn
ABSTRACT

Tandem duplication mutations are increasingly found to be the direct cause of many rare heritable diseases, accounting for up to 10% of cases. Unfortunately, animal models recapitulating such mutations are scarce, limiting our ability to study them and develop genome editing therapies. Here, we describe the generation of a novel duplication mouse model, harboring a multi-exonic tandem duplication in the Dmd gene which recapitulates a human mutation. Duplication correction of this mouse was achieved by implementing a single-guide RNA (sgRNA) CRISPR/Cas9 approach. This strategy precisely removed a duplication mutation in vivo, restored full-length dystrophin expression, and was accompanied by improvements in both histopathological and clinical phenotypes. We conclude that CRISPR/Cas9 represents a powerful tool to accurately model and treat tandem duplication mutations. Our findings will open new avenues of research for exploring the study and therapeutics of duplication disorders.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Insulin solution from bovine pancreas, BioReagent, 10 mg/mL insulin in 25  mM HEPES, pH 8.2, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Monoclonal Anti-Dystrophin antibody produced in mouse, clone MANDYS8, ascites fluid
Sigma-Aldrich
Tyrode′s Solution, Acidic, liquid, sterile-filtered, suitable for mouse embryo cell culture
Sigma-Aldrich
ESGRO® Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.