Generation of a gene-corrected human isogenic iPSC line from a patient with Fabry disease carrying the GLA variant c.1069C>T using CRISPR/Cas9-mediated homology directed repair

Fabry disease (FD) is an X-linked genetic disorder caused by mutations in the GLA gene, leading to α-galactosidase A deficiency and intracellular globotriaosylceramide (Gb3) accumulation. To study FD-associated pathomechanisms, we generated an isogenic control induced pluripotent stem cell (iPSC) li...

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Main Authors: Franziska Karl-Schöller, Maximilian Breyer, Eva Klopocki, Nurcan Üçeyler
Format: Article
Language:English
Published: Elsevier 2025-08-01
Series:Stem Cell Research
Online Access:http://www.sciencedirect.com/science/article/pii/S1873506125000613
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author Franziska Karl-Schöller
Maximilian Breyer
Eva Klopocki
Nurcan Üçeyler
author_facet Franziska Karl-Schöller
Maximilian Breyer
Eva Klopocki
Nurcan Üçeyler
author_sort Franziska Karl-Schöller
collection DOAJ
description Fabry disease (FD) is an X-linked genetic disorder caused by mutations in the GLA gene, leading to α-galactosidase A deficiency and intracellular globotriaosylceramide (Gb3) accumulation. To study FD-associated pathomechanisms, we generated an isogenic control induced pluripotent stem cell (iPSC) line (IsoFD-1) from a patient-derived FD-iPSC line (FD-1) carrying the GLA c.1069C>T mutation. Using CRISPR/Cas9 gene correction, we restored the wild-type sequence, confirmed by Sanger sequencing and absence of Gb3 deposits. IsoFD-1 exhibited typical pluripotency markers, normal karyotype, and trilineage differentiation capacity. This line provides a valuable tool for investigating Gb3-related cellular dysfunction in FD.
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spelling doaj-art-94599dec186a46bcb9dd7f0b4f77cfaf2025-08-20T03:10:46ZengElsevierStem Cell Research1873-50612025-08-018610371110.1016/j.scr.2025.103711Generation of a gene-corrected human isogenic iPSC line from a patient with Fabry disease carrying the GLA variant c.1069C>T using CRISPR/Cas9-mediated homology directed repairFranziska Karl-Schöller0Maximilian Breyer1Eva Klopocki2Nurcan Üçeyler3University Hospital Würzburg, Department of Neurology, 97080 Würzburg, GermanyUniversity Hospital Würzburg, Department of Neurology, 97080 Würzburg, GermanyUniversity of Würzburg, Institute for Human Genetics, 97074 Würzburg, GermanyUniversity Hospital Würzburg, Department of Neurology, 97080 Würzburg, Germany; Corresponding author.Fabry disease (FD) is an X-linked genetic disorder caused by mutations in the GLA gene, leading to α-galactosidase A deficiency and intracellular globotriaosylceramide (Gb3) accumulation. To study FD-associated pathomechanisms, we generated an isogenic control induced pluripotent stem cell (iPSC) line (IsoFD-1) from a patient-derived FD-iPSC line (FD-1) carrying the GLA c.1069C>T mutation. Using CRISPR/Cas9 gene correction, we restored the wild-type sequence, confirmed by Sanger sequencing and absence of Gb3 deposits. IsoFD-1 exhibited typical pluripotency markers, normal karyotype, and trilineage differentiation capacity. This line provides a valuable tool for investigating Gb3-related cellular dysfunction in FD.http://www.sciencedirect.com/science/article/pii/S1873506125000613
spellingShingle Franziska Karl-Schöller
Maximilian Breyer
Eva Klopocki
Nurcan Üçeyler
Generation of a gene-corrected human isogenic iPSC line from a patient with Fabry disease carrying the GLA variant c.1069C>T using CRISPR/Cas9-mediated homology directed repair
Stem Cell Research
title Generation of a gene-corrected human isogenic iPSC line from a patient with Fabry disease carrying the GLA variant c.1069C>T using CRISPR/Cas9-mediated homology directed repair
title_full Generation of a gene-corrected human isogenic iPSC line from a patient with Fabry disease carrying the GLA variant c.1069C>T using CRISPR/Cas9-mediated homology directed repair
title_fullStr Generation of a gene-corrected human isogenic iPSC line from a patient with Fabry disease carrying the GLA variant c.1069C>T using CRISPR/Cas9-mediated homology directed repair
title_full_unstemmed Generation of a gene-corrected human isogenic iPSC line from a patient with Fabry disease carrying the GLA variant c.1069C>T using CRISPR/Cas9-mediated homology directed repair
title_short Generation of a gene-corrected human isogenic iPSC line from a patient with Fabry disease carrying the GLA variant c.1069C>T using CRISPR/Cas9-mediated homology directed repair
title_sort generation of a gene corrected human isogenic ipsc line from a patient with fabry disease carrying the gla variant c 1069c t using crispr cas9 mediated homology directed repair
url http://www.sciencedirect.com/science/article/pii/S1873506125000613
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