{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T20:17:41Z","timestamp":1788985061953,"version":"build-2803163510"},"posted":{"date-parts":[[2026,7,6]]},"group-title":"Developmental Biology","reference-count":64,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T00:00:00Z","timestamp":1783296000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["NIH ORIP R24OD020166"],"award-info":[{"award-number":["NIH ORIP R24OD020166"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["NIH ORIP R24OD036201"],"award-info":[{"award-number":["NIH ORIP R24OD036201"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["NIH NIDDK R01DK131162"],"award-info":[{"award-number":["NIH NIDDK R01DK131162"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Zebrafish combines the power of genetics and unparalleled\n                  <jats:italic>in vivo<\/jats:italic>\n                  imaging to investigate the dynamics of vertebrate hematopoietic development. The conserved transcription factor Runx1 is essential for definitive hematopoiesis. We generated an endogenous zebrafish\n                  <jats:italic>runx1-2A-creERT2<\/jats:italic>\n                  CRISPR knock-in for tamoxifen-regulated Runx1 lineage tracing and characterized its activity using recombinase-dependent fluorescence reporters, live imaging and flow cytometry. Tamoxifen induction of Cre in the early embryo labeled all expected Runx1 lineages, including neuroectodermal olfactory placode and Rohan-Beard neurons, primitive hematopoietic blood cells, and emerging definitive hematopoietic stem and progenitor cells (HSPCs) in the dorsal aorta. Short windows of tamoxifen treatment allowed separation of Runx1 primitive from definitive hematopoiesis and visualization of HSPC colonization of the larval caudal hematopoietic tissue, thymus and kidney. Runx1 HSPCs labeled during development persisted into adulthood and contributed to hematopoietic precursors, myeloid, lymphoid, and peripheral blood lineages. Labeling of all blood lineages was also effective by tamoxifen treatment of 5-month-old adults. The\n                  <jats:italic>runx1-2A-creERT2<\/jats:italic>\n                  knock-in line provides a novel genetic tool for inducible spatial and temporal analysis of Runx1 progenitor mechanisms in developmental and adult hematopoiesis.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Summary Statement<\/jats:title>\n                  <jats:p>\n                    Zebrafish\n                    <jats:italic>runx1-2A-creERT2<\/jats:italic>\n                    knock-in provides inducible spatial and temporal Cre recombinase Runx1 lineage labeling in development and adults, and reveals embryonic Runx1 precursors contribute to adult hematopoiesis.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.07.03.736368","type":"posted-content","created":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T09:45:13Z","timestamp":1783331113000},"source":"Crossref","is-referenced-by-count":0,"title":["Tracing developmental and adult hematopoiesis with an endogenous zebrafish\n                  <i>runx1-2A- CreERT2<\/i>\n                  CRISPR knock-in"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3635-2297","authenticated-orcid":false,"given":"James A.","family":"Preston","sequence":"first","affiliation":[{"name":"Department of Genetics, Development and Cell Biology, Iowa State University, Ames, Iowa 50011- 1101 USA"},{"name":"Interdepartmental Genetics and Genomics Graduate Program, Iowa State University, Ames, Iowa 50011-1101 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-7635-9716","authenticated-orcid":false,"given":"Masuma K.","family":"Usha","sequence":"additional","affiliation":[{"name":"Department of Genetics, Development and Cell Biology, Iowa State University, Ames, Iowa 50011- 1101 USA"},{"name":"Interdepartmental Molecular, Cellular, and Developmental Biology Graduate Program, Iowa State University, Ames, Iowa 50011-1101 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0726-4212","authenticated-orcid":false,"given":"Stephen C.","family":"Ekker","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Department of Molecular Biosciences, University of Texas at Austin, Austin, Texas 78712 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9637-0967","authenticated-orcid":false,"given":"Karl J.","family":"Clark","sequence":"additional","affiliation":[{"name":"Department of Animal Science, Texas A&M University, College Station, Texas 77843-2471 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8816-3848","authenticated-orcid":false,"given":"Jeffrey J.","family":"Essner","sequence":"additional","affiliation":[{"name":"Department of Genetics, Development and Cell Biology, Iowa State University, Ames, Iowa 50011- 1101 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6033-5628","authenticated-orcid":false,"given":"Raquel","family":"Espin-Palazon","sequence":"additional","affiliation":[{"name":"Department of Genetics, Development and Cell Biology, Iowa State University, Ames, Iowa 50011- 1101 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9308-6189","authenticated-orcid":false,"given":"Maura","family":"McGrail","sequence":"additional","affiliation":[{"name":"Department of Genetics, Development and Cell Biology, Iowa State University, Ames, Iowa 50011- 1101 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026090913000673000_2026.07.03.736368v2.1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2005-01-0179"},{"key":"2026090913000673000_2026.07.03.736368v2.2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-11423-2"},{"key":"2026090913000673000_2026.07.03.736368v2.3","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2008-04-149898"},{"key":"2026090913000673000_2026.07.03.736368v2.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.12.032"},{"key":"2026090913000673000_2026.07.03.736368v2.5","doi-asserted-by":"publisher","DOI":"10.1038\/nature05883"},{"key":"2026090913000673000_2026.07.03.736368v2.6","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.64835"},{"key":"2026090913000673000_2026.07.03.736368v2.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2021.109703"},{"key":"2026090913000673000_2026.07.03.736368v2.8","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.53968"},{"key":"2026090913000673000_2026.07.03.736368v2.9","doi-asserted-by":"crossref","unstructured":"Welker, J.M. , Wierson, W.A. , Almeida, M.P. , Mann, C.M. , Torrie, M.E. , Ekker, S.C. , Clark, K.J. , Dobbs, D.L. , Essner, J.J. , McGrail, M . 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