{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T08:38:45Z","timestamp":1762072725520,"version":"build-2065373602"},"reference-count":66,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,10,20]],"date-time":"2022-10-20T00:00:00Z","timestamp":1666224000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deakin University"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>JAK3 is principally activated by members of the interleukin-2 receptor family and plays an essential role in lymphoid development, with inactivating JAK3 mutations causing autosomal-recessive severe combined immunodeficiency (SCID). This study aimed to generate an equivalent zebrafish model of SCID and to characterize the model across the life-course. Genome editing of zebrafish jak3 created mutants similar to those observed in human SCID. Homozygous jak3 mutants showed reduced embryonic T lymphopoiesis that continued through the larval stage and into adulthood, with B cell maturation and adult NK cells also reduced and neutrophils impacted. Mutant fish were susceptible to lymphoid leukemia. This model has many of the hallmarks of human SCID resulting from inactivating JAK3 mutations and will be useful for a variety of pre-clinical applications.<\/jats:p>","DOI":"10.3390\/biom12101521","type":"journal-article","created":{"date-parts":[[2022,10,20]],"date-time":"2022-10-20T20:35:55Z","timestamp":1666298155000},"page":"1521","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Zebrafish Model of Severe Combined Immunodeficiency (SCID) Due to JAK3 Mutation"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4850-2205","authenticated-orcid":false,"given":"Faiza","family":"Basheer","sequence":"first","affiliation":[{"name":"School of Medicine, Deakin University, Waurn Ponds, Geelong, VIC 3216, Australia"},{"name":"Institute of Mental and Physical Health and Clinical Translation, Deakin University, Waurn Ponds, Geelong, VIC 3216, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Effie","family":"Lee","sequence":"additional","affiliation":[{"name":"Gribbles Veterinary Pathology, Glenside, SA 5065, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1646-4905","authenticated-orcid":false,"given":"Clifford","family":"Liongue","sequence":"additional","affiliation":[{"name":"School of Medicine, Deakin University, Waurn Ponds, Geelong, VIC 3216, Australia"},{"name":"Institute of Mental and Physical Health and Clinical Translation, Deakin University, Waurn Ponds, Geelong, VIC 3216, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alister C.","family":"Ward","sequence":"additional","affiliation":[{"name":"School of Medicine, Deakin University, Waurn Ponds, Geelong, VIC 3216, Australia"},{"name":"Institute of Mental and Physical Health and Clinical Translation, Deakin University, Waurn Ponds, Geelong, VIC 3216, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2004433","DOI":"10.1002\/advs.202004433","article-title":"Cytokines: From clinical significance to quantification","volume":"8","author":"Liu","year":"2021","journal-title":"Adv. Sci."},{"key":"ref_2","first-page":"121","article-title":"Cytokines and their role in health and disease: A brief overview","volume":"4","author":"Gulati","year":"2016","journal-title":"Mol. Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Liongue, C., O\u2019Sullivan, L.A., Trengove, M.C., and Ward, A.C. (2012). Evolution of JAK-STAT pathway components: Mechanisms and role in immune system development. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0032777"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1186\/gb-2004-5-12-253","article-title":"The Janus kinases (Jaks)","volume":"5","author":"Yamaoka","year":"2004","journal-title":"Genome Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1111\/j.1600-065X.2008.00754.x","article-title":"Janus kinases in immune cell signaling","volume":"228","author":"Ghoreschi","year":"2009","journal-title":"Immunol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"28527","DOI":"10.1074\/jbc.270.48.28527","article-title":"Interleukins 2, 4, 7, and 15 stimulate tyrosine phosphorylation of insulin receptor substrates 1 and 2 in T cells potential role of Jak kinases","volume":"270","author":"Johnston","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1038\/370151a0","article-title":"Phosphorylation and activation of the Jak-3 Janus kinase in response to interleukin-2","volume":"370","author":"Johnston","year":"1994","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1878","DOI":"10.1073\/pnas.94.5.1878","article-title":"Requirement for an initial signal from the membrane-proximal region of the interleukin 2 receptor \u03b3c chain for Janus kinase activation leading to T cell proliferation","volume":"94","author":"Nelson","year":"1997","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1042","DOI":"10.1126\/science.7973658","article-title":"Interaction of IL-2R\u03b2 and \u03b3c chains with Jak1 and Jak3: Implications for XSCID and XCID","volume":"266","author":"Russell","year":"1994","journal-title":"Science"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2144","DOI":"10.1038\/leu.2012.74","article-title":"Newly described activating JAK3 mutations in T-cell acute lymphoblastic leukemia","volume":"26","author":"Bains","year":"2012","journal-title":"Leukemia"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1038\/sj.leu.2404527","article-title":"JAK3 mutations occur in acute megakaryoblastic leukemia both in Down syndrome children and non-Down syndrome adults","volume":"21","author":"Kiyoi","year":"2007","journal-title":"Leukemia"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1158\/2159-8290.CD-12-0028","article-title":"Janus kinase 3\u2013activating mutations identified in Natural killer\/T-cell lymphoma","volume":"2","author":"Koo","year":"2012","journal-title":"Cancer Discov."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1002\/humu.1188","article-title":"Mutations in severe combined immune deficiency (SCID) due to JAK3 deficiency","volume":"18","author":"Notarangelo","year":"2001","journal-title":"Hum. Mutat."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1038\/377065a0","article-title":"Mutations of Jak-3 gene in patients with autosomal severe combined immune deficiency (SCID)","volume":"377","author":"Macchi","year":"1995","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1126\/science.270.5237.797","article-title":"Mutation of Jak3 in a patient with SCID: Essential role of Jak3 in lymphoid development","volume":"270","author":"Russell","year":"1995","journal-title":"Science"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1186\/s13045-015-0126-4","article-title":"Zebrafish as a model for leukemia and other hematopoietic disorders","volume":"8","author":"Rasighaemi","year":"2015","journal-title":"J. Hematol. Oncol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Liongue, C., and Ward, A.C. (2007). Evolution of class I cytokine receptors. BMC Evol. Biol., 7.","DOI":"10.1186\/1471-2148-7-120"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Sertori, R., Jones, R., Basheer, F., Rivera, L., Dawson, S., Loke, S., Heidary, S., Dhillon, A., Liongue, C., and Ward, A.C. (2022). Generation and characterization of a zebrafish IL-2R\u03b3c SCID model. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23042385"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2575","DOI":"10.1084\/jem.20160378","article-title":"Single-cell imaging of normal and malignant cell engraftment into optically clear prkdc-null SCID zebrafish","volume":"213","author":"Moore","year":"2016","journal-title":"J. Exp. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"7060","DOI":"10.4049\/jimmunol.1003907","article-title":"Genetic evidence for an evolutionarily conserved role of IL-7 signaling in T cell development of zebrafish","volume":"186","author":"Iwanami","year":"2011","journal-title":"J. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1007\/s00018-022-04361-8","article-title":"In vivo impact of JAK3 A573V mutation revealed using zebrafish","volume":"79","author":"Basheer","year":"2022","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"887","DOI":"10.1126\/science.1080280","article-title":"Myc-induced T cell leukemia in transgenic zebrafish","volume":"299","author":"Langenau","year":"2003","journal-title":"Science"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.aquaculture.2007.04.077","article-title":"The husbandry of zebrafish (Danio rerio): A review","volume":"269","author":"Lawrence","year":"2007","journal-title":"Aquaculture"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Garritano, S., Gemignani, F., Voegele, C., Nguyen-Dumont, T., Calvez-Kelm, L., De Silva, D., Lesueur, F., Landi, S., and Tavtigian, S.V. (2009). Determining the effectiveness of High Resolution Melting analysis for SNP genotyping and mutation scanning at the TP53 locus. BMC Genet., 10.","DOI":"10.1186\/1471-2156-10-5"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1038\/nprot.2007.514","article-title":"High-resolution in situ hybridization to whole-mount zebrafish embryos","volume":"3","author":"Thisse","year":"2008","journal-title":"Nat. Protoc."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212 \u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2009","DOI":"10.1182\/blood-2003-06-2104","article-title":"Janus kinase 3 (JAK3) deficiency: Clinical, immunologic, and molecular analyses of 10 patients and outcomes of stem cell transplantation","volume":"103","author":"Roberts","year":"2004","journal-title":"Blood"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1002\/dvdy.1223","article-title":"Ikaros expression as a marker for lymphoid progenitors during zebrafish development","volume":"222","author":"Willett","year":"2001","journal-title":"Dev. Dyn."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1016\/j.dci.2003.12.003","article-title":"T cells and the thymus in developing zebrafish","volume":"28","author":"Danilova","year":"2004","journal-title":"Dev. Comp. Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7369","DOI":"10.1073\/pnas.0402248101","article-title":"In vivo tracking of T cell development, ablation, and engraftment in transgenic zebrafish","volume":"101","author":"Langenau","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/S0925-4773(01)00658-X","article-title":"Cloning and expression pattern of the lysozyme C gene in zebrafish","volume":"113","author":"Liu","year":"2002","journal-title":"Mech. Dev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3087","DOI":"10.1182\/blood.V98.10.3087","article-title":"Morphologic and functional characterization of granulocytes and macrophages in embryonic and adult zebrafish","volume":"98","author":"Lieschke","year":"2001","journal-title":"Blood"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1182\/blood-2012-12-471029","article-title":"An evolutionarily conserved program of B-cell development and activation in zebrafish","volume":"122","author":"Page","year":"2013","journal-title":"Blood"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Petrie-Hanson, L., Hohn, C., and Hanson, L. (2009). Characterization of rag 1 mutant zebrafish leukocytes. BMC Immunol., 10.","DOI":"10.1186\/1471-2172-10-8"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2463","DOI":"10.4049\/jimmunol.177.4.2463","article-title":"Conserved functions of Ikaros in vertebrate lymphocyte development: Genetic evidence for distinct larval and adult phases of T cell development and two lineages of B cells in zebrafish","volume":"177","author":"Schorpp","year":"2006","journal-title":"J. Immunol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1084\/jem.20152013","article-title":"Single-cell transcriptional analysis of normal, aberrant, and malignant hematopoiesis in zebrafish","volume":"213","author":"Moore","year":"2016","journal-title":"J. Exp. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1182\/blood.V89.2.688","article-title":"Characterization of adult \u03b1-and \u03b2-globin genes in the zebrafish","volume":"89","author":"Chan","year":"1997","journal-title":"Blood"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1182\/blood-2010-01-262873","article-title":"Macrophage-specific gene functions in Spi1-directed innate immunity","volume":"116","author":"Zakrzewska","year":"2010","journal-title":"Blood"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.immuni.2013.01.004","article-title":"Interleukin-2 at the crossroads of effector responses, tolerance, and immunotherapy","volume":"38","author":"Liao","year":"2013","journal-title":"Immunity"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1146\/annurev.immunol.26.021607.090357","article-title":"The biology of interleukin-2","volume":"26","author":"Malek","year":"2008","journal-title":"Annu. Rev. Immunol."},{"key":"ref_41","first-page":"861920","article-title":"Role of common-gamma chain cytokines in NK cell development and function: Perspectives for immunotherapy","volume":"2011","author":"Meazza","year":"2011","journal-title":"J. Biomed. Biotechnol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1038\/nri2580","article-title":"New insights into the regulation of T cells by \u03b3c family cytokines","volume":"9","author":"Rochman","year":"2009","journal-title":"Nat. Rev. Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2376","DOI":"10.1016\/j.biocel.2009.09.004","article-title":"JAK3: A two-faced player in hematological disorders","volume":"41","author":"Cornejo","year":"2009","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"800","DOI":"10.1126\/science.270.5237.800","article-title":"Defective lymphoid development in mice lacking Jak3","volume":"270","author":"Nosaka","year":"1995","journal-title":"Science"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1016\/1074-7613(95)90066-7","article-title":"Developmental defects of lymphoid cells in Jak3 kinase-deficient mice","volume":"3","author":"Park","year":"1995","journal-title":"Immunity"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1016\/S0952-7915(97)80108-2","article-title":"The role of Jak3 in lymphoid development, activation, and signaling","volume":"9","author":"Thomis","year":"1997","journal-title":"Curr. Opin. Immunol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"3151","DOI":"10.1182\/blood.V87.8.3151.bloodjournal8783151","article-title":"Murine JAK3 is preferentially expressed in hematopoietic tissues and lymphocyte precursor cells","volume":"87","author":"Gurniak","year":"1996","journal-title":"Blood"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"25028","DOI":"10.1074\/jbc.270.42.25028","article-title":"A kinase-deficient splice variant of the human JAK3 is expressed in hematopoietic and epithelial cancer cells","volume":"270","author":"Lai","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1093\/emboj\/18.6.1549","article-title":"Autosomal SCID caused by a point mutation in the N-terminus of Jak3: Mapping of the Jak3\u2013receptor interaction domain","volume":"18","author":"Cacalano","year":"1999","journal-title":"EMBO J."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3996","DOI":"10.1182\/blood.V90.10.3996","article-title":"Structural and functional basis for JAK3-deficient severe combined immunodeficiency","volume":"90","author":"Candotti","year":"1997","journal-title":"Blood"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/0092-8674(93)90167-O","article-title":"Interleukin-2 receptor \u03b3 chain mutation results in X-linked severe combined immunodeficiency in humans","volume":"73","author":"Noguchi","year":"1993","journal-title":"Cell"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"794","DOI":"10.1126\/science.270.5237.794","article-title":"Defects in B lymphocyte maturation and T lymphocyte activation in mice lacking Jak3","volume":"270","author":"Thomis","year":"1995","journal-title":"Science"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"401","DOI":"10.3389\/fgene.2019.00401","article-title":"Biological characteristics of severe combined immunodeficient mice produced by CRISPR\/Cas9-mediated Rag2 and IL2rg mutation","volume":"10","author":"Zhao","year":"2019","journal-title":"Front. Genet."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1538\/expanim.19-0120","article-title":"Generation of novel Il2rg-knockout mice with clustered regularly interspaced short palindromic repeats (CRISPR) and Cas9","volume":"69","author":"Byambaa","year":"2020","journal-title":"Exp. Anim."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"135","DOI":"10.4049\/jimmunol.1403060","article-title":"Conserved IL-2R\u03b3c signaling mediates lymphopoiesis in zebrafish","volume":"196","author":"Sertori","year":"2016","journal-title":"J. Immunol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1038\/3877","article-title":"Defective IL7R expression in T-B+ NK+ severe combined immunodeficiency","volume":"20","author":"Puel","year":"1998","journal-title":"Nat. Genet."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2803","DOI":"10.1182\/blood.V96.8.2803","article-title":"A partial deficiency of interleukin-7R\u03b1 is sufficient to abrogate T-cell development and cause severe combined immunodeficiency","volume":"96","author":"Roifman","year":"2000","journal-title":"Blood"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1955","DOI":"10.1084\/jem.180.5.1955","article-title":"Early lymphocyte expansion is severely impaired in interleukin 7 receptor-deficient mice","volume":"180","author":"Peschon","year":"1994","journal-title":"J. Exp. Med."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1519","DOI":"10.1084\/jem.181.4.1519","article-title":"Lymphopenia in interleukin (IL)-7 gene-deleted mice identifies IL-7 as a nonredundant cytokine","volume":"181","author":"Vieira","year":"1995","journal-title":"J. Exp. Med."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1084\/jem.184.1.289","article-title":"Interleukin-7 receptor alpha is essential for the development of gamma delta+ T cells, but not Natural killer cells","volume":"184","author":"He","year":"1996","journal-title":"J. Exp. Med."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1182\/blood.V94.3.932.415k30_932_939","article-title":"Dysregulated myelopoiesis in mice lacking Jak3","volume":"94","author":"Grossman","year":"1999","journal-title":"Blood"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"2753","DOI":"10.1182\/blood.V100.8.2753","article-title":"Activation of the Jak3 pathway is associated with granulocytic differentiation of myeloid precursor cells","volume":"100","author":"Rane","year":"2002","journal-title":"Blood"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1016\/j.molimm.2004.04.014","article-title":"Jak3 and the pathogenesis of severe combined immunodeficiency","volume":"41","author":"Husa","year":"2004","journal-title":"Mol. Immunol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/S0145-305X(03)00103-4","article-title":"Development and maturation of the immune system in zebrafish, Danio rerio: A gene expression profiling, in situ hybridization and immunological study","volume":"28","author":"Lam","year":"2004","journal-title":"Dev. Comp. Immunol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1038\/ni.2549","article-title":"Fishing for mammalian paradigms in the teleost immune system","volume":"14","author":"Sunyer","year":"2013","journal-title":"Nat. Immunol."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"1028","DOI":"10.1016\/j.jaci.2017.05.024","article-title":"Cancer in primary immunodeficiency diseases: Cancer incidence in the United States Immune Deficiency Network Registry","volume":"141","author":"Mayor","year":"2018","journal-title":"J. Allergy Clin. Immunol."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/12\/10\/1521\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:57:56Z","timestamp":1760144276000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/12\/10\/1521"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,20]]},"references-count":66,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["biom12101521"],"URL":"https:\/\/doi.org\/10.3390\/biom12101521","relation":{},"ISSN":["2218-273X"],"issn-type":[{"type":"electronic","value":"2218-273X"}],"subject":[],"published":{"date-parts":[[2022,10,20]]}}}