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A core erythroid transcriptional network, together with non-DNA-binding cofactors, occupies regulatory regions of genes essential for erythroid development. This process is further shaped by epigenetic mechanisms, including histone post-translational modifications and long-range chromatin interactions. CCCTC-binding factor (CTCF) is a multifunctional regulator with a central role in three-dimensional chromatin organization. Although CTCF has been implicated in hematopoietic differentiation and leukemogenesis, its specific function in erythropoiesis remains poorly defined. Here, we investigated the role of CTCF during erythroid differentiation using two complementary models: pluripotent K562 leukemia cells and primary human CD34+ hematopoietic stem\/progenitor cells, each induced toward the erythroid lineage by distinct stimuli. In both systems, CTCF silencing impaired erythroid differentiation by repression of key erythroid transcription factor genes, including LMO2, KLF1, MYB, and ETS1. This repression was associated with enrichment of repressive histone marks at CTCF-binding sites within their regulatory regions. Moreover, CTCF cooperated with cohesin to establish and stabilize long-range chromatin interactions at these loci. These results provide new insight into how CTCF-dependent chromatin regulation contributes to normal erythroid development and suggest that perturbation of this regulatory axis may have implications for hematopoietic disorders and malignancies.<\/jats:p>","DOI":"10.3390\/biom16040549","type":"journal-article","created":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T09:49:44Z","timestamp":1775641784000},"page":"549","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["CTCF Regulates Erythroid Differentiation Through Control of Core Erythroid Transcription Factors"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5998-0344","authenticated-orcid":false,"given":"Lorena","family":"Garc\u00eda-Gaipo","sequence":"first","affiliation":[{"name":"Institute of Biomedicine and Biotechnology of Cantabria (IBBTEC), CSIC\u2013University of Cantabria, 39011 Santander, Spain"},{"name":"Department of Molecular Biology, University of Cantabria, 39011 Santander, Spain"},{"name":"Cell Cycle, Stem Cell Fate and Cancer Laboratory, Marqu\u00e9s de Valdecilla Research Institute (IDIVAL), 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5782-6370","authenticated-orcid":false,"given":"Vanessa","family":"Junco","sequence":"additional","affiliation":[{"name":"Institute of Biomedicine and Biotechnology of Cantabria (IBBTEC), CSIC\u2013University of Cantabria, 39011 Santander, Spain"},{"name":"Department of Molecular Biology, University of Cantabria, 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4396-4814","authenticated-orcid":false,"given":"Luc\u00eda","family":"Garc\u00eda-Guti\u00e9rrez","sequence":"additional","affiliation":[{"name":"Institute of Biomedicine and Biotechnology of Cantabria (IBBTEC), CSIC\u2013University of Cantabria, 39011 Santander, Spain"},{"name":"Department of Molecular Biology, University of Cantabria, 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ver\u00f3nica","family":"Torrano","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of the Basque Country (UPV\/EHU), 48940 Leioa, Spain"},{"name":"Biomedical Research Networking Center on Cancer (CIBERONC), 28029 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosa","family":"Blanco","sequence":"additional","affiliation":[{"name":"Institute of Biomedicine and Biotechnology of Cantabria (IBBTEC), CSIC\u2013University of Cantabria, 39011 Santander, Spain"},{"name":"Department of Molecular Biology, University of Cantabria, 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandra","family":"Wiesinger","sequence":"additional","affiliation":[{"name":"Institute of Biomedicine and Biotechnology of Cantabria (IBBTEC), CSIC\u2013University of Cantabria, 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-2175-6024","authenticated-orcid":false,"given":"Rujula","family":"Pradeep","sequence":"additional","affiliation":[{"name":"Department of Data Science, The Fu Foundation School of Engineering and Applied Science, Columbia University, New York, NY 10027, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose Luis","family":"Arroyo","sequence":"additional","affiliation":[{"name":"Blood and Tissue Bank of Cantabria, Marqu\u00e9s de Valdecilla Foundation, Marqu\u00e9s de Valdecilla Research Institute (IDIVAL), 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Batlle-L\u00f3pez","sequence":"additional","affiliation":[{"name":"Institute of Biomedicine and Biotechnology of Cantabria (IBBTEC), CSIC\u2013University of Cantabria, 39011 Santander, Spain"},{"name":"Department of Hematology, Marqu\u00e9s de Valdecilla University Hospital\u2013IDIVAL, 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5803-0112","authenticated-orcid":false,"given":"Javier","family":"Le\u00f3n","sequence":"additional","affiliation":[{"name":"Institute of Biomedicine and Biotechnology of Cantabria (IBBTEC), CSIC\u2013University of Cantabria, 39011 Santander, Spain"},{"name":"Department of Molecular Biology, University of Cantabria, 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2169-3972","authenticated-orcid":false,"given":"Manuel","family":"Rosa-Garrido","sequence":"additional","affiliation":[{"name":"Institute of Biomedicine and Biotechnology of Cantabria (IBBTEC), CSIC\u2013University of Cantabria, 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4682-4040","authenticated-orcid":false,"given":"M. Dolores","family":"Delgado","sequence":"additional","affiliation":[{"name":"Institute of Biomedicine and Biotechnology of Cantabria (IBBTEC), CSIC\u2013University of Cantabria, 39011 Santander, Spain"},{"name":"Department of Molecular Biology, University of Cantabria, 39011 Santander, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2099","DOI":"10.1038\/s12276-024-01311-1","article-title":"Erythropoiesis: Insights from a genomic perspective","volume":"56","author":"Cha","year":"2024","journal-title":"Exp. Mol. 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