{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:11:56Z","timestamp":1792789916572,"version":"4.5.0"},"reference-count":58,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T00:00:00Z","timestamp":1670889600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Clinician Scientist Program","award":["N\u00b0. 440-0-0"],"award-info":[{"award-number":["N\u00b0. 440-0-0"]}]},{"name":"Stefan Morsch Stiftung, MaxCyte Inc.","award":["N\u00b0. 440-0-0"],"award-info":[{"award-number":["N\u00b0. 440-0-0"]}]},{"name":"the University Children\u2019s Hospital of T\u00fcbingen","award":["N\u00b0. 440-0-0"],"award-info":[{"award-number":["N\u00b0. 440-0-0"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>X-linked severe combined immunodeficiency (X-SCID) is a primary immunodeficiency that is caused by mutations in the interleukin-2 receptor gamma (IL2RG) gene. Some patients present atypical X-SCID with mild clinical symptoms due to somatic revertant mosaicism. CRISPR\/Cas9 and prime editing are two advanced genome editing tools that paved the way for treating immune deficiency diseases. Prime editing overcomes the limitations of the CRISPR\/Cas9 system, as it does not need to induce double-strand breaks (DSBs) or exogenous donor DNA templates to modify the genome. Here, we applied CRISPR\/Cas9 with single-stranded oligodeoxynucleotides (ssODNs) and prime editing methods to generate an in vitro model of the disease in K\u2013562 cells and healthy donors\u2019 T cells for the c. 458T&gt;C point mutation in the IL2RG gene, which also resulted in a useful way to optimize the gene correction approach for subsequent experiments in patients\u2019 cells. Both methods proved to be successful and were able to induce the mutation of up to 31% of treated K\u2013562 cells and 26% of treated T cells. We also applied similar strategies to correct the IL2RG c. 458T&gt;C mutation in patient T cells that carry the mutation with revertant somatic mosaicism. However, both methods failed to increase the frequency of the wild-type sequence in the mosaic T cells of patients due to limited in vitro proliferation of mutant cells and the presence of somatic reversion. To the best of our knowledge, this is the first attempt to treat mosaic cells from atypical X-SCID patients employing CRISPR\/Cas9 and prime editing. We showed that prime editing can be applied to the formation of specific-point IL2RG mutations without inducing nonspecific on-target modifications. We hypothesize that the feasibility of the nucleotide substitution of the IL2RG gene using gene therapy, especially prime editing, could provide an alternative strategy to treat X-SCID patients without revertant mutations, and further technological improvements need to be developed to correct somatic mosaicism mutations.<\/jats:p>","DOI":"10.3390\/genes13122348","type":"journal-article","created":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T03:32:32Z","timestamp":1670902352000},"page":"2348","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Challenges in Gene Therapy for Somatic Reverted Mosaicism in X-Linked Combined Immunodeficiency by CRISPR\/Cas9 and Prime Editing"],"prefix":"10.3390","volume":"13","author":[{"given":"Yujuan","family":"Hou","sequence":"first","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4699-7486","authenticated-orcid":false,"given":"Guillermo","family":"Ure\u00f1a-Bail\u00e9n","sequence":"additional","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tahereh","family":"Mohammadian Gol","sequence":"additional","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul Gerhard","family":"Gratz","sequence":"additional","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hans Peter","family":"Gratz","sequence":"additional","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0775-5820","authenticated-orcid":false,"given":"Alicia","family":"Roig-Merino","sequence":"additional","affiliation":[{"name":"MaxCyte Inc., Rockville, MD 20850, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justin S.","family":"Antony","sequence":"additional","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4464-0152","authenticated-orcid":false,"given":"Andr\u00e9s","family":"Lamsfus-Calle","sequence":"additional","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alberto","family":"Daniel-Moreno","sequence":"additional","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rupert","family":"Handgretinger","sequence":"additional","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0211-2343","authenticated-orcid":false,"given":"Markus","family":"Mezger","sequence":"additional","affiliation":[{"name":"Department of Pediatrics I, Hematology and Oncology, University Children\u2019s Hospital, University of T\u00fcbingen, 72076 Tuebingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.1093\/hmg\/2.8.1099","article-title":"The interleukin-2 receptor \u03b3 chain maps to Xq13.1 and is mutated in X-linked severe combined immunodeficiency, SCIDX1","volume":"2","author":"Puck","year":"1993","journal-title":"Hum. Mol. Genet."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3749","DOI":"10.1182\/blood-2013-02-380105","article-title":"How I treat severe combined immunodeficiency","volume":"122","author":"Gaspar","year":"2013","journal-title":"Blood"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.gendis.2019.07.004","article-title":"Genetics of severe combined immunodeficiency","volume":"7","author":"Kumrah","year":"2020","journal-title":"Genes Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s13223-018-0317-y","article-title":"hypomorphic mutation: Identification of a novel pathogenic mutation in exon 8 and a review of the literature","volume":"15","author":"Lim","year":"2019","journal-title":"Allergy Asthma Clin. Immunol. Off. J. Can. Soc. Allergy Clin. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1563","DOI":"10.1056\/NEJM199611213352104","article-title":"Atypical X-linked severe combined immunodeficiency due to possible spontaneous reversion of the genetic defect in T cells","volume":"335","author":"Stephan","year":"1996","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1007\/s10875-012-9684-1","article-title":"Multiple reversions of an IL2RG mutation restore T cell function in an X-linked severe combined immunodeficiency patient","volume":"32","author":"Kawai","year":"2012","journal-title":"J. Clin. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1007\/s10875-015-0174-0","article-title":"IL2RG reversion event in a common lymphoid progenitor leads to delayed diagnosis and milder phenotype","volume":"35","author":"Hsu","year":"2015","journal-title":"J. Clin. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1030","DOI":"10.3324\/haematol.2012.077511","article-title":"A reversion of an IL2RG mutation in combined immunodeficiency providing competitive advantage to the majority of CD8+ T cells","volume":"98","author":"Kuijpers","year":"2013","journal-title":"Haematologica"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"102961","DOI":"10.1016\/j.ebiom.2020.102961","article-title":"Long-term robustness of a T-cell system emerging from somatic rescue of a genetic block in T-cell development","volume":"59","author":"Kury","year":"2020","journal-title":"EBioMedicine"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1007\/s10875-015-0202-0","article-title":"Late-Onset Combined Immunodeficiency with a Novel IL2RG Mutation and Probable Revertant Somatic Mosaicism","volume":"35","author":"Okuno","year":"2015","journal-title":"J. Clin. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"4090","DOI":"10.1182\/blood-2008-04-153361","article-title":"Clinical and immunologic consequences of a somatic reversion in a patient with X-linked severe combined immunodeficiency","volume":"112","author":"Speckmann","year":"2008","journal-title":"Blood"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1007\/s10875-020-00825-3","article-title":"Progressive B Cell Loss in Revertant X-SCID","volume":"40","author":"Lin","year":"2020","journal-title":"J. Clin. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Hou, Y., Gratz, H.P., Ure\u00f1a-Bail\u00e9n, G., Gratz, P.G., Schilbach-St\u00fcckle, K., Renno, T., G\u00fcng\u00f6r, D., Mader, D.A., Malenke, E., and Antony, J.S. (2021). Somatic Reversion of a Novel Mutation Resulting in Atypical X-Linked Combined Immunodeficiency. Genes, 13.","DOI":"10.3390\/genes13010035"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/S0022-3476(97)70199-9","article-title":"Human severe combined immunodeficiency: Genetic, phenotypic, and functional diversity in one hundred eight infants","volume":"130","author":"Buckley","year":"1997","journal-title":"J. Pediatr."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"608653","DOI":"10.3389\/fimmu.2020.608653","article-title":"Immune Reconstitution After Gene Therapy Approaches in Patients With X-Linked Severe Combined Immunodeficiency Disease","volume":"11","author":"Blanco","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1038\/sj.mt.6300393","article-title":"Self-inactivating gammaretroviral vectors for gene therapy of X-linked severe combined immunodeficiency","volume":"16","author":"Thornhill","year":"2008","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"718","DOI":"10.1038\/mt.2008.5","article-title":"Physiological promoters reduce the genotoxic risk of integrating gene vectors","volume":"16","author":"Zychlinski","year":"2008","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1016\/j.jaci.2020.05.055","article-title":"Gene therapy for X-linked severe combined immunodeficiency: Historical outcomes and current status","volume":"146","author":"Pai","year":"2020","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"R15","DOI":"10.1093\/hmg\/ddz170","article-title":"Gene therapy for primary immunodeficiency","volume":"28","author":"Booth","year":"2019","journal-title":"Hum. Mol. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1634","DOI":"10.1038\/s41467-019-09614-y","article-title":"Gene correction for SCID-X1 in long-term hematopoietic stem cells","volume":"10","author":"Wiebking","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Rozov, S.M., Permyakova, N.V., and Deineko, E.V. (2019). The Problem of the Low Rates of CRISPR\/Cas9-Mediated Knock-ins in Plants: Approaches and Solutions. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20133371"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1038\/s41586-019-1711-4","article-title":"Search-and-replace genome editing without double-strand breaks or donor DNA","volume":"576","author":"Anzalone","year":"2019","journal-title":"Nature"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1445","DOI":"10.1089\/hum.2019.29098.trf","article-title":"Prime Editing: A Novel Cas9-Reverse Transcriptase Fusion May Revolutionize Genome Editing","volume":"30","author":"Flotte","year":"2019","journal-title":"Hum. Gene Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1073\/pnas.97.1.274","article-title":"Diversity, functionality, and stability of the T cell repertoire derived in vivo from a single human T cell precursor","volume":"97","author":"Bousso","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"W171","DOI":"10.1093\/nar\/gkz365","article-title":"CHOPCHOP v3: Expanding the CRISPR web toolbox beyond genome editing","volume":"47","author":"Labun","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"570","DOI":"10.1038\/mt.2015.197","article-title":"Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR\/Cas9, and megaTAL Nucleases","volume":"24","author":"Osborn","year":"2016","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1038\/s41551-020-00622-8","article-title":"A web tool for the design of prime-editing guide RNAs","volume":"5","author":"Chow","year":"2021","journal-title":"Nat. Biomed. Eng."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"10133","DOI":"10.1038\/s41598-020-66309-x","article-title":"Comparative targeting analysis of KLF1, BCL11A, and HBG1\/2 in CD34 HSPCs by CRISPR\/Cas9 for the induction of fetal hemoglobin","volume":"10","author":"Antony","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1089\/crispr.2021.0075","article-title":"A Mutation-Agnostic Hematopoietic Stem Cell Gene Therapy for Metachromatic Leukodystrophy","volume":"5","author":"Antony","year":"2022","journal-title":"CRISPR J."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1089\/crispr.2021.0113","article-title":"Inference of CRISPR Edits from Sanger Trace Data","volume":"5","author":"Conant","year":"2022","journal-title":"CRISPR J."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1038\/nature17664","article-title":"Efficient introduction of specific homozygous and heterozygous mutations using CRISPR\/Cas9","volume":"533","author":"Paquet","year":"2016","journal-title":"Nature"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"iyab199","DOI":"10.1093\/genetics\/iyab199","article-title":"Single nucleotide substitutions effectively block Cas9 and allow for scarless genome editing in Caenorhabditis elegans","volume":"220","author":"Medley","year":"2022","journal-title":"Genetics"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.1016\/j.ymthe.2018.02.028","article-title":"Nuclease-free Adeno-Associated Virus-Mediated Il2rg Gene Editing in X-SCID Mice","volume":"26","author":"Hiramoto","year":"2018","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1126\/science.1088547","article-title":"LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1","volume":"302","author":"Schmidt","year":"2003","journal-title":"Science"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1056\/NEJM200301163480314","article-title":"A serious adverse event after successful gene therapy for X-linked severe combined immunodeficiency","volume":"348","author":"Schmidt","year":"2003","journal-title":"N. Engl. J. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3132","DOI":"10.1172\/JCI35700","article-title":"Insertional oncogenesis in 4 patients after retrovirus-mediated gene therapy of SCID-X1","volume":"118","author":"Garrigue","year":"2008","journal-title":"J. Clin. Investig."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1525","DOI":"10.1056\/NEJMoa1815408","article-title":"Lentiviral Gene Therapy Combined with Low-Dose Busulfan in Infants with SCID-X1","volume":"380","author":"Mamcarz","year":"2019","journal-title":"N. Engl. J. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1407","DOI":"10.1056\/NEJMoa1404588","article-title":"A modified \u03b3-retrovirus vector for X-linked severe combined immunodeficiency","volume":"371","author":"Pai","year":"2014","journal-title":"N. Engl. J. Med."},{"key":"ref_39","first-page":"335ra357","article-title":"Lentiviral hematopoietic stem cell gene therapy for X-linked severe combined immunodeficiency","volume":"8","author":"Wu","year":"2016","journal-title":"Sci. Transl. Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1038\/nrg2842","article-title":"Genome editing with engineered zinc finger nucleases","volume":"11","author":"Urnov","year":"2010","journal-title":"Nat. Rev. Genet."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"eaan0820","DOI":"10.1126\/scitranslmed.aan0820","article-title":"Preclinical modeling highlights the therapeutic potential of hematopoietic stem cell gene editing for correction of SCID-X1","volume":"9","author":"Schiroli","year":"2017","journal-title":"Sci. Transl. Med."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1038\/nature03556","article-title":"Highly efficient endogenous human gene correction using designed zinc-finger nucleases","volume":"435","author":"Urnov","year":"2005","journal-title":"Nature"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1298","DOI":"10.1038\/nbt1353","article-title":"Gene editing in human stem cells using zinc finger nucleases and integrase-defective lentiviral vector delivery","volume":"25","author":"Lombardo","year":"2007","journal-title":"Nat. Biotechnol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1038\/nrm3486","article-title":"TALENs: A widely applicable technology for targeted genome editing","volume":"14","author":"Joung","year":"2013","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1016\/j.stem.2015.02.005","article-title":"Lymphoid regeneration from gene-corrected SCID-X1 subject-derived iPSCs","volume":"16","author":"Menon","year":"2015","journal-title":"Cell Stem Cell"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1038\/nbt.2842","article-title":"CRISPR-Cas systems for editing, regulating and targeting genomes","volume":"32","author":"Sander","year":"2014","journal-title":"Nat. Biotechnol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1038\/nature13420","article-title":"Targeted genome editing in human repopulating haematopoietic stem cells","volume":"510","author":"Genovese","year":"2014","journal-title":"Nature"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1186\/s13287-021-02565-6","article-title":"Homology-directed gene-editing approaches for hematopoietic stem and progenitor cell gene therapy","volume":"12","author":"Azhagiri","year":"2021","journal-title":"Stem Cell Res. Ther."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1262","DOI":"10.1016\/j.cell.2014.05.010","article-title":"Development and applications of CRISPR-Cas9 for genome engineering","volume":"157","author":"Hsu","year":"2014","journal-title":"Cell"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1126\/science.1231143","article-title":"Multiplex genome engineering using CRISPR\/Cas systems","volume":"339","author":"Cong","year":"2013","journal-title":"Science"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"4811","DOI":"10.1038\/s41598-019-41121-4","article-title":"Highly efficient genome editing for single-base substitutions using optimized ssODNs with Cas9-RNPs","volume":"9","author":"Okamoto","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/s40348-018-0086-1","article-title":"Gene correction of HBB mutations in CD34+ hematopoietic stem cells using Cas9 mRNA and ssODN donors","volume":"5","author":"Antony","year":"2018","journal-title":"Mol. Cell. Pediatr."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"e79208","DOI":"10.7554\/eLife.79208","article-title":"Highly efficient generation of isogenic pluripotent stem cell models using prime editing","volume":"11","author":"Li","year":"2022","journal-title":"eLife"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"425","DOI":"10.2353\/jmoldx.2010.090188","article-title":"Comparison of Sanger sequencing, pyrosequencing, and melting curve analysis for the detection of KRAS mutations: Diagnostic and clinical implications","volume":"12","author":"Tsiatis","year":"2010","journal-title":"J. Mol. Diagn. JMD"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"783022","DOI":"10.3389\/fimmu.2021.783022","article-title":"Reversion Mosaicism in Primary Immunodeficiency Diseases","volume":"12","author":"Miyazawa","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"5635","DOI":"10.1016\/j.cell.2021.09.018","article-title":"Enhanced prime editing systems by manipulating cellular determinants of editing outcomes","volume":"184","author":"Chen","year":"2021","journal-title":"Cell"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1038\/nrg3424","article-title":"A genomic view of mosaicism and human disease","volume":"14","author":"Biesecker","year":"2013","journal-title":"Nat. Rev. Genet."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"718","DOI":"10.1007\/s10875-021-01037-z","article-title":"Genetic Mosaicism as a Cause of Inborn Errors of Immunity","volume":"41","author":"Aluri","year":"2021","journal-title":"J. Clin. Immunol."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/13\/12\/2348\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:40:16Z","timestamp":1760146816000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/13\/12\/2348"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,13]]},"references-count":58,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["genes13122348"],"URL":"https:\/\/doi.org\/10.3390\/genes13122348","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,13]]}}}