{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,23]],"date-time":"2026-03-23T01:00:30Z","timestamp":1774227630524,"version":"3.50.1"},"reference-count":42,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2025,2,28]],"date-time":"2025-02-28T00:00:00Z","timestamp":1740700800000},"content-version":"vor","delay-in-days":58,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"European Union\u2019s Horizon Europe"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,1,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>CRISPR\/Cas9-mediated homology-directed repair (HDR) allows precise gene editing, but its efficiency remains low for certain cell types, such as human induced pluripotent stem cells (hiPSCs). In this study, we aimed to introduce the CTNNA1: c.2023C&amp;gt;T (p.Q675*) genetic alteration, which is associated with Hereditary Diffuse Gastric Cancer, into hiPSCs using CRISPR\/Cas9. We designed a single-guide RNA targeting the alteration site and a single-stranded oligonucleotide donor DNA template for HDR-based repair. Herein, we report the successful introduction of the CTNNA1: c.2023C&amp;gt;T homozygous alteration in one hiPSC line, which resulted in severe phenotypic changes, including impaired colony formation and cell proliferation. Additionally, we established a straightforward protocol to assess hiPSCs karyotype integrity, ensuring the chromosomal stability required for the gene-editing process. This protocol involves routine G-banding analysis that is required for regular quality controls during handling of hiPSCs. This study demonstrates an efficient approach to precisely edit hiPSCs by CRISPR\/Cas9 and highlights the essential role of CTNNA1 expression in maintaining hiPSC viability. Our methodology provides a valuable framework for modeling disease-associated alterations in human-derived cellular models that can be reproduced for other genes and other types of cell lines.<\/jats:p>","DOI":"10.1093\/biomethods\/bpaf018","type":"journal-article","created":{"date-parts":[[2025,3,4]],"date-time":"2025-03-04T21:50:13Z","timestamp":1741125013000},"source":"Crossref","is-referenced-by-count":4,"title":["A protocol for karyotyping and genetic editing of induced pluripotent stem cells with homology-directed repair mediated CRISPR\/Cas9"],"prefix":"10.1093","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5705-2958","authenticated-orcid":false,"given":"Silvana","family":"Lobo","sequence":"first","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, University of Porto , Porto, 4200-135,","place":["Portugal"]},{"name":"ICBAS\u2014Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar, University of Porto , Porto, 4050-313,","place":["Portugal"]}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0794-4912","authenticated-orcid":false,"given":"Rita","family":"Barbosa-Matos","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, University of Porto , Porto, 4200-135,","place":["Portugal"]}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9225-9076","authenticated-orcid":false,"given":"Sofia","family":"D\u00f3ria","sequence":"additional","affiliation":[{"name":"Genetics Unit, Department of Pathology, Faculty of Medicine, University of Porto , Porto, 4200-319,","place":["Portugal"]}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-1159-3621","authenticated-orcid":false,"given":"Ana Maria","family":"Pedro","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, University of Porto , Porto, 4200-135,","place":["Portugal"]}]},{"given":"Ana","family":"Brito","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, University of Porto , Porto, 4200-135,","place":["Portugal"]}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9712-1460","authenticated-orcid":false,"given":"Daniel","family":"Ferreira","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, University of Porto , Porto, 4200-135,","place":["Portugal"]}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8340-2264","authenticated-orcid":false,"given":"Carla","family":"Oliveira","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, University of Porto , Porto, 4200-135,","place":["Portugal"]},{"name":"IPATIMUP\u2014Institute of Molecular Pathology and Immunology of the University of Porto , Porto, 4200-135,","place":["Portugal"]},{"name":"FMUP\u2014Faculdade de Medicina, University of Porto , Porto, 4200-319,","place":["Portugal"]}]}],"member":"286","published-online":{"date-parts":[[2025,2,28]]},"reference":[{"key":"2025032320514377000_bpaf018-B1","doi-asserted-by":"publisher","first-page":"1258096","DOI":"10.1126\/science.1258096","article-title":"Genome editing. 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