Functional Correction of Dystrophin Actin Binding Domain Mutations by Genome Editing

Overview

Journal JCI Insight

Specialties Biomedical Engineering
General Medicine

Date 2017 Sep 22

PMID 28931764

Citations 57

Authors

Viktoriia Kyrychenko

Sergii Kyrychenko

Malte Tiburcy

John M Shelton

Chengzu Long

Jay W Schneider

Wolfram-Hubertus Zimmermann

Rhonda Bassel-Duby

Eric N Olson

Affiliations

Soon will be listed here.

Abstract

Dystrophin maintains the integrity of striated muscles by linking the actin cytoskeleton with the cell membrane. Duchenne muscular dystrophy (DMD) is caused by mutations in the dystrophin gene (DMD) that result in progressive, debilitating muscle weakness, cardiomyopathy, and a shortened lifespan. Mutations of dystrophin that disrupt the amino-terminal actin-binding domain 1 (ABD-1), encoded by exons 2-8, represent the second-most common cause of DMD. In the present study, we compared three different strategies for CRISPR/Cas9 genome editing to correct mutations in the ABD-1 region of the DMD gene by deleting exons 3-9, 6-9, or 7-11 in human induced pluripotent stem cells (iPSCs) and by assessing the function of iPSC-derived cardiomyocytes. All three exon deletion strategies enabled the expression of truncated dystrophin protein and restoration of cardiomyocyte contractility and calcium transients to varying degrees. We show that deletion of exons 3-9 by genomic editing provides an especially effective means of correcting disease-causing ABD-1 mutations. These findings represent an important step toward eventual correction of common DMD mutations and provide a means of rapidly assessing the expression and function of internally truncated forms of dystrophin-lacking portions of ABD-1.

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