1. Need for high-resolution genetic analysis in iPSC: results and lessons from the ForIPS consortium
- Author
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Martin Regensburger, Mandy Krumbiegel, Steffen Uebe, Sonja Ploetz, Anita Rauch, Udo Trautmann, Arif B. Ekici, Frank Edenhofer, Katharina Günther, Annika Sommer, Beate Winner, Bernt Popp, Cornelia Kraus, Zacharias Kohl, André Reis, Michaela Farrell, Janina Grosch, Jürgen Winkler, Reza Asadollahi, University of Zurich, and Reis, André
- Subjects
0301 basic medicine ,10039 Institute of Medical Genetics ,Induced Pluripotent Stem Cells ,Clone (cell biology) ,Cell Culture Techniques ,lcsh:Medicine ,610 Medicine & health ,Computational biology ,Biology ,Genetic analysis ,Germline ,Article ,03 medical and health sciences ,Humans ,Copy-number variation ,Allele ,lcsh:Science ,Induced pluripotent stem cell ,QH426 ,Exome sequencing ,Biological Specimen Banks ,1000 Multidisciplinary ,Multidisciplinary ,lcsh:R ,High-Throughput Nucleotide Sequencing ,Sequence Analysis, DNA ,Genetic Profile ,DNA Fingerprinting ,030104 developmental biology ,Karyotyping ,570 Life sciences ,biology ,lcsh:Q ,Reprogramming - Abstract
Genetic integrity of induced pluripotent stem cells (iPSCs) is essential for their validity as disease models and for potential therapeutic use. We describe the comprehensive analysis in the ForIPS consortium: an iPSC collection from donors with neurological diseases and healthy controls. Characterization included pluripotency confirmation, fingerprinting, conventional and molecular karyotyping in all lines. In the majority, somatic copy number variants (CNVs) were identified. A subset with available matched donor DNA was selected for comparative exome sequencing. We identified single nucleotide variants (SNVs) at different allelic frequencies in each clone with high variability in mutational load. Low frequencies of variants in parental fibroblasts highlight the importance of germline samples. Somatic variant number was independent from reprogramming, cell type and passage. Comparison with disease genes and prediction scores suggest biological relevance for some variants. We show that high-throughput sequencing has value beyond SNV detection and the requirement to individually evaluate each clone.
- Published
- 2018