CRISPR/Cas9 genome editing in LGMD2A/R1 patient-derived induced pluripotent stem and skeletal muscle progenitor cells

  • Large numbers of Calpain 3 (CAPN3) mutations cause recessive forms of limb-girdle muscular dystrophy (LGMD2A/LGMDR1) with selective atrophy of the proximal limb muscles. We have generated induced pluripotent stem cells (iPSC) from a patient with two mutations in exon 3 and exon 4 at the calpain 3 locus (W130C, 550delA). Two different strategies to rescue these mutations are devised: (i) on the level of LGMD2A-iPSC, we combined CRISPR/Cas9 genome targeting with a FACS and Tet transactivator-based biallelic selection strategy, which resulted in a new functional chimeric exon 3-4 without the two CAPN3 mutations. (ii) On the level of LGMD2A-iPSC-derived CD82+/Pax7+ myogenic progenitor cells, we demonstrate CRISPR/Cas9 mediated rescue of the highly prevalent exon 4 CAPN3 mutation. The first strategy specifically provides isogenic LGMD2A corrected iPSC for disease modelling, and the second strategy can be further elaborated for potential translational approaches.

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Author:Lampros MavrommatisGND, Abdul ZabenGND, Urs KindlerORCiDGND, Marie-Cécile Wellner-KienitzGND, Julienne DietzGND, Hyun-Woo JeongGND, Pierre BöhmeGND, Beate Brand-SaberiORCiDGND, Matthias VorgerdORCiDGND, Holm ZaehresORCiDGND
Parent Title (English):Stem cells international
Publisher:Hindawi Publishing Corporation, part of Wiley
Place of publication:London
Document Type:Article
Date of Publication (online):2024/04/15
Date of first Publication:2023/11/09
Publishing Institution:Ruhr-Universität Bochum, Universitätsbibliothek
Tag:Open Access Fonds
Issue:Artikel 9246825
First Page:9246825-1
Last Page:9246825-17
Article Processing Charge funded by the Deutsche Forschungsgemeinschaft (DFG) and the Open Access Publication Fund of Ruhr-Universität Bochum.
Institutes/Facilities:Institut für Anatomie, Abteilung für Anatomie und Molekulare Embryologie
Dewey Decimal Classification:Technik, Medizin, angewandte Wissenschaften / Medizin, Gesundheit
open_access (DINI-Set):open_access
faculties:Medizinische Fakultät
Licence (English):License LogoCreative Commons - CC BY 4.0 - Attribution 4.0 International