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  • In vivo non-invasive monitoring of dystrophin correction in a new Duchenne muscular dystrophy reporter mouse.

In vivo non-invasive monitoring of dystrophin correction in a new Duchenne muscular dystrophy reporter mouse.

Nature communications (2019-10-06)
Leonela Amoasii, Hui Li, Yu Zhang, Yi-Li Min, Efrain Sanchez-Ortiz, John M Shelton, Chengzu Long, Alex A Mireault, Samadrita Bhattacharyya, John R McAnally, Rhonda Bassel-Duby, Eric N Olson
ABSTRACT

Duchenne muscular dystrophy (DMD) is a fatal genetic disorder caused by mutations in the dystrophin gene. To enable the non-invasive analysis of DMD gene correction strategies in vivo, we introduced a luciferase reporter in-frame with the C-terminus of the dystrophin gene in mice. Expression of this reporter mimics endogenous dystrophin expression and DMD mutations that disrupt the dystrophin open reading frame extinguish luciferase expression. We evaluated the correction of the dystrophin reading frame coupled to luciferase in mice lacking exon 50, a common mutational hotspot, after delivery of CRISPR/Cas9 gene editing machinery with adeno-associated virus. Bioluminescence monitoring revealed efficient and rapid restoration of dystrophin protein expression in affected skeletal muscles and the heart. Our results provide a sensitive non-invasive means of monitoring dystrophin correction in mouse models of DMD and offer a platform for testing different strategies for amelioration of DMD pathogenesis.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal Anti-Vinculin antibody produced in mouse, clone hVIN-1, ascites fluid
Sigma-Aldrich
Anti-Cas9 Antibody, clone 7A9, clone 7A9, from mouse
Sigma-Aldrich
Monoclonal Anti-Dystrophin antibody produced in mouse, clone MANDYS8, ascites fluid