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Genet."],"published-print":{"date-parts":[[2026,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Human induced pluripotent stem cells (hiPSCs) represent a powerful platform for disease modeling, especially in monogenic diseases as they preserve the donor\u2019s genetic background while enabling directed differentiation into disease-relevant cell types. This makes them highly suitable for studying disease mechanisms in a patient-specific and physiologically relevant context. Although CRISPR\/Cas9 is widely applied for genome editing, precise correction of pathogenic variants in hiPSCs remains challenging due to the lack of standardized CRISPR component selection and experimental design. Here, we describe an optimized CRISPR-based strategy for correcting a heterozygous\n                    <jats:italic>HNF1A<\/jats:italic>\n                    frameshift mutation (c.235_236insG; p.Glu79Glyfs*16) in HNF1A-MODY patient-derived hiPSCs. Using electroporation, we efficiently delivered CRISPR components, including a ribonucleoprotein complex of Cas9 and single-guide RNA, along with a single-stranded oligodeoxynucleotide repair template. Corrected hiPSC lines were validated for pluripotency, absence of exogenous reprogramming factors, and off-target effects. Additionally, we discuss key technical challenges encountered during the editing process and provide practical recommendations that may improve the generation of mutation-corrected hiPSC lines. These guidelines could serve as a useful reference for researchers employing CRISPR-based strategies for generation of reliable disease modelling tools.\n                  <\/jats:p>","DOI":"10.1007\/s00439-026-02863-0","type":"journal-article","created":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T08:02:52Z","timestamp":1786521772000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["CRISPR\/Cas9-based repair of a heterozygous HNF1A mutation in patient-derived hiPSCs"],"prefix":"10.1007","volume":"145","author":[{"given":"Dawid","family":"Skoczek","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jerzy","family":"Hohendorff","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maciej T.","family":"Malecki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alicia","family":"Roig-Merino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7383-0297","authenticated-orcid":false,"given":"Rasmus O.","family":"Bak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3226-0726","authenticated-orcid":false,"given":"Neli","family":"Kachamakova-Trojanowska","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,12]]},"reference":[{"key":"2863_CR1","doi-asserted-by":"publisher","first-page":"140","DOI":"10.1186\/s40478-024-01851-7","volume":"12","author":"K Ahuja","year":"2024","unstructured":"Ahuja K, Vandenabeele M, Nami F et al (2024) A deep phenotyping study in mouse and iPSC models to understand the role of oligodendroglia in optic neuropathy in Wolfram syndrome. 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ROB holds equity in Kamau Therapeutics and UNIKUM Therapeutics and is a cofounder of UNIKUM Therapeutics. Neither company was involved in the present study. ROB is an inventor on patents and patent applications related to CRISPR\/Cas and cellular therapies. ROB reports research funding from Novo Nordisk. All other authors do not have competing interests.","order":1,"name":"Ethics","label":"Conflict of interest","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The blood collection for preparation of the patient-specific hiPSCs was done after obtaining the informed consent of the patient and following the approval of the Jagiellonian University Bioethical Committee (No. 1072.6120.310.2018).","order":2,"name":"Ethics","label":"Ethical approval","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Figures\n                      \n                      and\n                      \n                      \u00a0A were created using Microsoft Office PowerPoint. Figure\n                      \n                      A was generated using the R programming environment. Figures\n                      \n                      B, S5 (gel electrophoresis), and 4B (PVDF membrane) are based on images captured using the ChemiDoc system (Bio-Rad). Figures\u00a0\n                      \n                      B and S2 include Sanger sequencing chromatograms visualized with SnapGene Viewer (Insightful Science; available at\n                      \n                      ). Figure\u00a0\n                      \n                      includes fluorescence microscopy images acquired using LAS X software (Leica Microsystems). Figure\u00a0\n                      \n                      A and C were prepared with GraphPad software. Figure\u00a0\n                      \n                      was created with BioRender (\n                      \n                      ). All figures (1\u20135) were assembled using Microsoft Office PowerPoint.","order":3,"name":"Ethics","label":"Figure preparation","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"69"}}