{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T14:51:10Z","timestamp":1785250270457,"version":"3.55.0"},"reference-count":19,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2024,5,28]],"date-time":"2024-05-28T00:00:00Z","timestamp":1716854400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deutsche Forschungsgemeinschaft (DFG)","award":["KU3389\/1-1"],"award-info":[{"award-number":["KU3389\/1-1"]}]},{"name":"Deutsche Forschungsgemeinschaft (DFG)","award":["SCHN1166\/4-1"],"award-info":[{"award-number":["SCHN1166\/4-1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Chronic granulomatous disease (CGD) is an inherited immunodeficiency disease mainly caused by mutations in the X-linked CYBB gene that abrogate reactive oxygen species (ROS) production in phagocytes and microbial defense. Gene repair using the CRISPR\/Cas9 system in hematopoietic stem and progenitor cells (HSPCs) is a promising technology for therapy for CGD. To support the establishment of efficient and safe gene therapies for CGD, we generated a mouse model harboring a patient-derived mutation in the CYBB gene. Our CybbC517del mouse line shows the hallmarks of CGD and provides a source for Cybb-deficient HSPCs that can be used to evaluate gene-therapy approaches in vitro and in vivo. In a setup using Cas9 RNPs and an AAV repair vector in HSPCs, we show that the mutation can be repaired in 19% of treated cells and that treatment restores ROS production by macrophages. In conclusion, our CybbC517del mouse line provides a new platform for refining and evaluating novel gene therapies and studying X-CGD pathophysiology.<\/jats:p>","DOI":"10.3390\/genes15060706","type":"journal-article","created":{"date-parts":[[2024,5,28]],"date-time":"2024-05-28T07:38:39Z","timestamp":1716881919000},"page":"706","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["A Mouse Model of X-Linked Chronic Granulomatous Disease for the Development of CRISPR\/Cas9 Gene Therapy"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-2771-4149","authenticated-orcid":false,"given":"Seren","family":"Sevim-Wunderlich","sequence":"first","affiliation":[{"name":"Max-Delbr\u00fcck-Center for Molecular Medicine in the Helmholtz Association (MDC), 13125 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tu","family":"Dang","sequence":"additional","affiliation":[{"name":"Max-Delbr\u00fcck-Center for Molecular Medicine in the Helmholtz Association (MDC), 13125 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jana","family":"Rossius","sequence":"additional","affiliation":[{"name":"Max-Delbr\u00fcck-Center for Molecular Medicine in the Helmholtz Association (MDC), 13125 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2426-6685","authenticated-orcid":false,"given":"Frank","family":"Schn\u00fctgen","sequence":"additional","affiliation":[{"name":"Department of Medicine, Hematology\/Oncology, University Hospital Frankfurt, Goethe University, 60590 Frankfurt am Main, Germany"},{"name":"German Cancer Consortium (DKTK), Partner Site Frankfurt\/Mainz, and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany"},{"name":"Frankfurt Cancer Institute, Goethe-University Frankfurt, 60596 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1694-9803","authenticated-orcid":false,"given":"Ralf","family":"K\u00fchn","sequence":"additional","affiliation":[{"name":"Max-Delbr\u00fcck-Center for Molecular Medicine in the Helmholtz Association (MDC), 13125 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"van den Berg, J.M., van Koppen, E., \u00c5hlin, A., Belohradsky, B.H., Bernatowska, E., Corbeel, L., Espan\u00f1ol, T., Fischer, A., Kurenko-Deptuch, M., and Mouy, R. (2009). Chronic Granulomatous Disease: The European Experience. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0005234"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1038\/ni.1992","article-title":"Germline CYBB Mutations That Selectively Affect Macrophages in Kindreds with X-Linked Predisposition to Tuberculous Mycobacterial Disease","volume":"12","author":"Bustamante","year":"2011","journal-title":"Nat. Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1663","DOI":"10.1182\/blood.V87.5.1663.1663","article-title":"Mutations in the X-Linked and Autosomal Recessive Forms of Chronic Granulomatous Disease","volume":"87","author":"Roos","year":"1996","journal-title":"Blood"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1016\/j.clim.2008.07.022","article-title":"X-Linked Chronic Granulomatous Disease Secondary to Skewed X Chromosome Inactivation in a Female with a Novel CYBB Mutation and Late Presentation","volume":"129","author":"Lewis","year":"2008","journal-title":"Clin. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1002\/humu.10212","article-title":"Human Gene Mutation Database (HGMD): 2003 Update","volume":"21","author":"Stenson","year":"2003","journal-title":"Hum. Mutat."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1200","DOI":"10.1002\/humu.20405","article-title":"Immunodeficiency Mutation Databases (IDbases)","volume":"27","author":"Vihinen","year":"2006","journal-title":"Hum. Mutat."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1517\/14712598.4.9.1423","article-title":"Gene Therapy for Chronic Granulomatous Disease","volume":"4","author":"Barese","year":"2004","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1097\/00005792-200005000-00004","article-title":"Genetic, Biochemical, and Clinical Features of Chronic Granulomatous Disease","volume":"79","author":"Segal","year":"2000","journal-title":"Medicine"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1007\/s00281-008-0121-8","article-title":"Genetics and Immunopathology of Chronic Granulomatous Disease","volume":"30","author":"Stasia","year":"2008","journal-title":"Semin. Immunopathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"eaah3480","DOI":"10.1126\/scitranslmed.aah3480","article-title":"CRISPR-Cas9 Gene Repair of Hematopoietic Stem Cells from Patients with X-Linked Chronic Granulomatous Disease","volume":"9","author":"Li","year":"2017","journal-title":"Sci. Transl. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1016\/j.bcmd.2010.07.012","article-title":"Hematologically Important Mutations: X-Linked Chronic Granulomatous Disease (Third Update)","volume":"45","author":"Roos","year":"2010","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_12","first-page":"207","article-title":"Gene Editing in Mouse Zygotes Using the CRISPR\/Cas9 System. Methods Mol","volume":"2631","author":"Wefers","year":"2023","journal-title":"Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1038\/s41551-019-0501-5","article-title":"Cytosine and Adenine Base Editing of the Brain, Liver, Retina, Heart and Skeletal Muscle of Mice via Adeno-Associated Viruses","volume":"4","author":"Levy","year":"2020","journal-title":"Nat. Biomed. Eng."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1038\/s41586-020-03086-7","article-title":"In Vivo Base Editing Rescues Hutchinson-Gilford Progeria Syndrome in Mice","volume":"589","author":"Koblan","year":"2021","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"eabm9106","DOI":"10.1126\/sciadv.abm9106","article-title":"Precise CRISPR-Cas-Mediated Gene Repair with Minimal off-Target and Unintended on-Target Mutations in Human Hematopoietic Stem Cells","volume":"8","author":"Tran","year":"2022","journal-title":"Sci. Adv."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1061","DOI":"10.1172\/JCI90031","article-title":"Loss of MicroRNA-7a2 Induces Hypogonadotropic Hypogonadism and Infertility","volume":"127","author":"Ahmed","year":"2017","journal-title":"J. Clin. Investig."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/S0022-1759(98)00136-7","article-title":"A Simple Flow Cytometry Assay Using Dihydrorhodamine for the Measurement of the Neutrophil Respiratory Burst in Whole Blood: Comparison with the Quantitative Nitrobluetetrazolium Test","volume":"219","author":"Richardson","year":"1998","journal-title":"J. Immunol. Methods"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1038\/ng0295-202","article-title":"Mouse Model of X-Linked Chronic Granulomatous Disease, an Inherited Defect in Phagocyte Superoxide Production","volume":"9","author":"Pollock","year":"1995","journal-title":"Nat. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2598","DOI":"10.1182\/blood.2020008503","article-title":"Enhanced Homology-Directed Repair for Highly Efficient Gene Editing in Hematopoietic Stem\/Progenitor Cells","volume":"137","author":"Brault","year":"2021","journal-title":"Blood"}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/15\/6\/706\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:49:31Z","timestamp":1760107771000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/15\/6\/706"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,28]]},"references-count":19,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2024,6]]}},"alternative-id":["genes15060706"],"URL":"https:\/\/doi.org\/10.3390\/genes15060706","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,28]]}}}