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Precise correction of heterozygous SHOX2 mutations in hiPSCs derived from patients with atrial fibrillation via genome editing and sib selection

Patient-specific human induced pluripotent stem cells (hiPSCs) offer unprecedented opportunities for the investigation of multigenic disease, personalized medicine, and stem cell therapy. For heterogeneous diseases such as atrial fibrillation (AF), however, precise correction of the associated mutat...

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Main Authors: Sumer, Simon (Author) , Hoffmann, Sandra (Author) , Laue, Svenja (Author) , Campbell, Birgit (Author) , Rädecke, Kristin (Author) , Frajs, Viktoria (Author) , Clauss, Sebastian (Author) , Kääb, Stefan (Author) , Janssen, Johannes W. G. (Author) , Jauch, Anna (Author) , Laugwitz, Karl-Ludwig (Author) , Dorn, Tatjana (Author) , Moretti, Alessandra (Author) , Rappold, Gudrun (Author)
Format: Article (Journal)
Language:English
Published: September 24, 2020
In: Stem cell reports
Year: 2020, Volume: 15, Issue: 4, Pages: 999-1013
ISSN:2213-6711
DOI:10.1016/j.stemcr.2020.08.015
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.stemcr.2020.08.015
Verlag, lizenzpflichtig, Volltext: http://www.sciencedirect.com/science/article/pii/S2213671120303465
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Author Notes:Simon Alexander Sumer, Sandra Hoffmann, Svenja Laue, Birgit Campbell, Kristin Raedecke, Viktoria Frajs, Sebastian Clauss, Stefan Kääb, Johannes W.G. Janssen, Anna Jauch, Karl-Ludwig Laugwitz, Tatjana Dorn, Alessandra Moretti, and Gudrun A. Rappold
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Summary:Patient-specific human induced pluripotent stem cells (hiPSCs) offer unprecedented opportunities for the investigation of multigenic disease, personalized medicine, and stem cell therapy. For heterogeneous diseases such as atrial fibrillation (AF), however, precise correction of the associated mutation is crucial. Here, we generated and corrected hiPSC lines from two AF patients carrying different heterozygous SHOX2 mutations. We developed a strategy for the scarless correction of heterozygous mutations, based on stochastic enrichment by sib selection, followed by allele quantification via digital PCR and next-generation sequencing to detect isogenic subpopulations. This allowed enriching edited cells 8- to 20-fold. The method does not require antibiotic selection or cell sorting and can be easily combined with base-and-prime editing approaches. Our strategy helps to overcome low efficiencies of homology-dependent repair in hiPSCs and facilitates the generation of isogenic control lines that represent the gold standard for modeling complex diseases in vitro.
Item Description:Gesehen am 30.11.2020
Physical Description:Online Resource
ISSN:2213-6711
DOI:10.1016/j.stemcr.2020.08.015

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