Functional correction of dystrophin actin binding domain mutations by genome editing

Viktoriia Kyrychenko, Sergii Kyrychenko, Malte Tiburcy, John M. Shelton, Chengzu Long, Jay W Schneider, Wolfram Hubertus Zimmermann, Rhonda S Bassel-Duby, Eric N Olson

Research output: Contribution to journalArticle

25 Citations (Scopus)

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.

Original languageEnglish (US)
JournalJCI insight
Volume2
Issue number18
DOIs
StatePublished - Sep 21 2017

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Dystrophin
Duchenne Muscular Dystrophy
Actins
Exons
Mutation
Induced Pluripotent Stem Cells
Cardiac Myocytes
Clustered Regularly Interspaced Short Palindromic Repeats
Striated Muscle
Muscle Weakness
Actin Cytoskeleton
Cardiomyopathies
Genes
Cell Membrane
Gene Editing
Calcium
Proteins

Keywords

  • Gene therapy
  • Genetic diseases
  • Muscle
  • Muscle Biology

Cite this

Kyrychenko, V., Kyrychenko, S., Tiburcy, M., Shelton, J. M., Long, C., Schneider, J. W., ... Olson, E. N. (2017). Functional correction of dystrophin actin binding domain mutations by genome editing. JCI insight, 2(18). https://doi.org/10.1172/jci.insight.95918

Functional correction of dystrophin actin binding domain mutations by genome editing. / Kyrychenko, Viktoriia; Kyrychenko, Sergii; Tiburcy, Malte; Shelton, John M.; Long, Chengzu; Schneider, Jay W; Zimmermann, Wolfram Hubertus; Bassel-Duby, Rhonda S; Olson, Eric N.

In: JCI insight, Vol. 2, No. 18, 21.09.2017.

Research output: Contribution to journalArticle

Kyrychenko, V, Kyrychenko, S, Tiburcy, M, Shelton, JM, Long, C, Schneider, JW, Zimmermann, WH, Bassel-Duby, RS & Olson, EN 2017, 'Functional correction of dystrophin actin binding domain mutations by genome editing', JCI insight, vol. 2, no. 18. https://doi.org/10.1172/jci.insight.95918
Kyrychenko V, Kyrychenko S, Tiburcy M, Shelton JM, Long C, Schneider JW et al. Functional correction of dystrophin actin binding domain mutations by genome editing. JCI insight. 2017 Sep 21;2(18). https://doi.org/10.1172/jci.insight.95918
Kyrychenko, Viktoriia ; Kyrychenko, Sergii ; Tiburcy, Malte ; Shelton, John M. ; Long, Chengzu ; Schneider, Jay W ; Zimmermann, Wolfram Hubertus ; Bassel-Duby, Rhonda S ; Olson, Eric N. / Functional correction of dystrophin actin binding domain mutations by genome editing. In: JCI insight. 2017 ; Vol. 2, No. 18.
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