biostudies-literatureRomero-Moya D"la Caixa" Foundation (Caixa Foundation)Ministry of Economy and Competitiveness | Instituto de Salud Carlos III (Institute of Health Carlos III)EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 Marie Skłodowska-Curie Actions (H2020 Excellent Science - Marie Skłodowska-Curie Actions)European Hematology Association (EHA)Ministerio de Economía y Competitividad (Ministry of Economy and Competitiveness)Ministry of Economy and Competitiveness | Agencia Estatal de Investigación (Spanish Agencia Estatal de Investigación)3015-3025https://www.ebi.ac.uk/biostudies/studies/S-EPMC12634441biostudies-literatureUnknown39(12)GATA2 deficiency is a monogenic transcriptopathy disorder characterized by bone marrow failure (BMF), immunodeficiency, and a high risk of developing myelodysplastic neoplasms (MDS) and acute myeloid leukemia (AML). Although informative mouse models have been developed, the mechanisms by which GATA2 haploinsufficiency drives disease initiation in humans remain incompletely understood. To address this, we developed a novel humanized model using CRISPR/Cas9 technology to knock-in GATA2-R398W variant in primary cord blood CD34⁺ cells. Additionally, we introduced specific mutations in SETBP1 and ASXL1 to model distinct premalignant stages of GATA2 deficiency. Through clonal competition and serial transplantation assays, we demonstrated that human CD34⁺ cells harboring the GATA2 mutaLeukemiaCRISPR-engineered human GATA2 deficiency model uncovers mitotic dysfunction and premature aging in HSPCs, impairing hematopoietic fitness.PMC12634441PID2023-151556OB-I00LCF-PR-HR24-00150FORT23/00032PID2020-15591RB-100AC23_2/00040PID2022-142966OB-I00KOG-202109-01162AC23_2/00014101029927Iglesias ASchilling MWlodarski MWPera JCatala ADe Giorgio FCastano JRomero-Moya DBerenguer-Balaguer CBigas AGiorgetti AGonzalez JMolina OPlass MMarin-Bejar OCalvo CMagallon-Mosella MDistefano MTorralba-Sales EPasquali LfalseCRISPR-engineered human GATA2 deficiency model uncovers mitotic dysfunction and premature aging in HSPCs, impairing hematopoietic fitness.GATA2 deficiency is a monogenic transcriptopathy disorder characterized by bone marrow failure (BMF), immunodeficiency, and a high risk of developing myelodysplastic neoplasms (MDS) and acute myeloid leukemia (AML). Although informative mouse models have been developed, the mechanisms by which GATA2 haploinsufficiency drives disease initiation in humans remain incompletely understood. To address this, we developed a novel humanized model using CRISPR/Cas9 technology to knock-in GATA2-R398W variant in primary cord blood CD34⁺ cells. Additionally, we introduced specific mutations in SETBP1 and ASXL1 to model distinct premalignant stages of GATA2 deficiency. Through clonal competition and serial transplantation assays, we demonstrated that human CD34⁺ cells harboring the GATA2 muta2025-01-01T00:00:00Z2025 Dec2026-06-12T04:48:36.434Z2026-06-12T03:07:40.884ZS-EPMC126344414095421510.1038/s41375-025-02771-8