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In recent years, it has been established that epigenetics plays an essential role in these mechanisms via the regulation of chromatin structure. Bromodomain-containing factors (BDFs), known for recognizing acetylated lysines on histones, have emerged as key factors in chromatin remodeling complexes. Among the eight predicted BDFs in T. cruzi, BDF6 is part of a TINTIN-like complex with MRGx and MRGBP, homologous to components of the NuA4\/TIP60 chromatin remodeling complex. We generated knockout (KO) parasites for the bdf6 gene using CRISPR\/Cas9 gene editing. BDF6 KO epimastigotes exhibit normal morphology but decreased size and growth, and the resulting metacyclic trypomastigotes displayed drastically reduced infectivity. Strikingly, once inside host cells, KO parasites differentiated into amastigotes but failed to replicate. This intracellular arrest was reversed by episomal complementation of BDF6. Consistently, KO parasites also exhibited impaired infectivity in mice, a defect that was also rescued in the addback parasite lines. Our findings highlight BDF6 as a critical regulator of intracellular parasite development, operating in stages beyond epimastigotes where epigenetic plasticity is essential for host adaptation. This striking stage-specific phenotype of BDF6 KO underscores its functional importance and highlights the relevance of epigenetic regulators along T. cruzi life cycle.<\/jats:p>","DOI":"10.1021\/acsinfecdis.6c00417","type":"journal-article","created":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T04:02:17Z","timestamp":1790136137000},"page":"3430-3444","source":"Crossref","is-referenced-by-count":0,"title":["BDF6 Deficiency\nSeverely Compromises Intracellular\nAmastigote Development and Infectivity of\n                    <i>Trypanosoma cruzi<\/i>"],"prefix":"10.1021","volume":"12","author":[{"given":"Victoria","family":"Boselli Raposo","sequence":"first","affiliation":[{"name":"CONICET\u2212Universidad Nacional de Rosario , , ,","place":["Rosario, Argentina, S2002LRK"]},{"name":"Universidad Nacional de Rosario , , ,","place":["Rosario, Argentina, S2002LRK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Virginia","family":"Perdomo","sequence":"additional","affiliation":[{"name":"Universidad Nacional de Rosario , , ,","place":["Rosario, Argentina, S2002LRK"]},{"name":"Consejo Nacional de Investigaciones Cienti\u0301ficas y Te\u0301cnicas (CONICET) , , ,","place":["Rosario, Santa Fe, Argentina, 2000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Florencia","family":"Pacini","sequence":"additional","affiliation":[{"name":"CONICET\u2212Universidad Nacional de Rosario , , ,","place":["Rosario, Argentina, 2000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan A.","family":"Cueto","sequence":"additional","affiliation":[{"name":"CONICET\u2212Universidad Nacional de Cuyo , , , ,","place":["Mendoza, Mendoza, Argentina, 5500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro","family":"Pezza","sequence":"additional","affiliation":[{"name":"CONICET\u2212Universidad Nacional de Rosario , , ,","place":["Rosario, Argentina, S2002LRK"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia S.","family":"Romano","sequence":"additional","affiliation":[{"name":"CONICET\u2212Universidad Nacional de Cuyo , , , ,","place":["Mendoza, Mendoza, Argentina, 5500"]},{"name":"Universidad Nacional de Cuyo , , ,","place":["Mendoza, Argentina, 5500"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4229-8784","authenticated-orcid":true,"given":"Roberto","family":"Docampo","sequence":"additional","affiliation":[{"name":"University of Georgia , , , ,","place":["Athens, Georgia, United States, 30602-7399"]},{"name":"University of Georgia , , , ,","place":["Athens, Georgia, United States, 30602-2607"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Rosa","family":"Perez","sequence":"additional","affiliation":[{"name":"CONICET\u2212Universidad Nacional de Rosario , , ,","place":["Rosario, Argentina, 2000"]},{"name":"Universidad Nacional de Rosario , , ,","place":["Rosario, Argentina, 2000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5986-7459","authenticated-orcid":true,"given":"Esteban","family":"Serra","sequence":"additional","affiliation":[{"name":"CONICET\u2212Universidad Nacional de Rosario , , ,","place":["Rosario, Argentina, S2002LRK"]},{"name":"Universidad Nacional de Rosario , , ,","place":["Rosario, Argentina, S2002LRK"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,22]]},"reference":[{"key":"2026100909101416600_cit1","unstructured":"PAHO\/WHO.\n          . Chagas disease. https:\/\/www.paho.org\/en\/topics\/chagas-disease#3."},{"issue":"S7301","key":"2026100909101416600_cit2","doi-asserted-by":"publisher","first-page":"S6","DOI":"10.1038\/nature09221","article-title":"Chagas\nDisease: A New Worldwide Challenge","volume":"465","author":"Coura","year":"2010","journal-title":"Nature"},{"key":"2026100909101416600_cit3","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1016\/j.csbj.2016.12.002","article-title":"Chagas Disease\nResearch and Development: Is There Light at the End of the Tunnel?","volume":"15","author":"Chatelain","year":"2017","journal-title":"Comput. Struct. Biotechnol. J."},{"issue":"14","key":"2026100909101416600_cit4","doi-asserted-by":"publisher","first-page":"1671","DOI":"10.2174\/1381612826999201203213527","article-title":"Basic Biology of Trypanosoma Cruzi","volume":"27","author":"Zuma","year":"2021","journal-title":"Curr. Pharm. Des."},{"key":"2026100909101416600_cit5","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.8947","article-title":"Transcriptomic\nChanges across the Life Cycle of Trypanosoma Cruzi II","volume":"8","author":"Cruz-Saavedra","year":"2020","journal-title":"PeerJ"},{"issue":"1","key":"2026100909101416600_cit6","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1042\/bse0510031","article-title":"Gene Expression Regulation in Trypanosomatids","volume":"51","author":"De Gaudenzi","year":"2011","journal-title":"Essays Biochem."},{"issue":"6","key":"2026100909101416600_cit7","doi-asserted-by":"publisher","DOI":"10.1098\/rsob.190072","article-title":"Regulation\nof Gene Expression in Trypanosomatids: Living with Polycistronic Transcription","volume":"9","author":"Clayton","year":"2019","journal-title":"Open Biol."},{"issue":"10","key":"2026100909101416600_cit8","doi-asserted-by":"publisher","first-page":"1196","DOI":"10.3390\/genes11101196","article-title":"Trypanosoma Cruzi Genome: Organization, Multi-Gene\nFamilies, Transcription, and Biological Implications","volume":"11","author":"Herreros-Cabello","year":"2020","journal-title":"Genes"},{"issue":"5","key":"2026100909101416600_cit9","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01079-20","article-title":"Histone Modifications\nand Other Facets of Epigenetic Regulation in Trypanosomatids: Leaving\nTheir Mark","volume":"11","author":"Saha","year":"2020","journal-title":"mBio"},{"issue":"12","key":"2026100909101416600_cit10","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1010056","article-title":"Reading\u201d a New\nChapter in\nProtozoan Parasite Transcriptional Regulation","volume":"17","author":"Fleck","year":"2021","journal-title":"PLOS Pathog."},{"key":"2026100909101416600_cit11","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1155\/2010\/525241","article-title":"Gene Expression in Trypanosomatid Parasites","volume":"2010","author":"Marti\u0301nez-Calvillo","year":"2010","journal-title":"J. Biomed. Biotechnol."},{"key":"2026100909101416600_cit12","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2025.118377","article-title":"The Epigenetic Landscape\nof Kinetoplastid Parasites: From Histone Post-Translational Modifications\nto Emerging Therapeutic Strategies","volume":"131","author":"Jacquet","year":"2025","journal-title":"Bioorg.\nMed. Chem."},{"key":"2026100909101416600_cit13","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/978-3-319-18102-8_1","article-title":"The Bromodomain\nas the Acetyl-Lysine Binding Domain in Gene Transcription","volume-title":"Histone Recognition","author":"Smith","year":"2015"},{"issue":"5","key":"2026100909101416600_cit14","doi-asserted-by":"publisher","first-page":"728","DOI":"10.1016\/j.molcel.2014.05.016","article-title":"The Mechanisms behind the Therapeutic Activity of BET\nBromodomain Inhibition","volume":"54","author":"Shi","year":"2014","journal-title":"Mol. Cell"},{"issue":"5","key":"2026100909101416600_cit15","doi-asserted-by":"publisher","first-page":"1062","DOI":"10.1021\/acsinfecdis.2c00057","article-title":"Essential\nBromodomain Tc BDF2 as a Drug Target against Chagas\nDisease","volume":"8","author":"Pezza","year":"2022","journal-title":"ACS Infect. Dis."},{"issue":"8","key":"2026100909101416600_cit16","doi-asserted-by":"publisher","first-page":"1081","DOI":"10.1590\/0074-0276140238","article-title":"Construction of\nThree New Gateway Expression Plasmids for Trypanosoma Cruzi","volume":"109","author":"Alonso","year":"2014","journal-title":"Mem. Inst. Oswaldo Cruz"},{"issue":"6","key":"2026100909101416600_cit17","doi-asserted-by":"publisher","first-page":"1267","DOI":"10.1021\/acsinfecdis.3c00148","article-title":"Deciphering Divergent Trypanosomatid Nuclear Complexes by Analyzing\nInteractomic Datasets with AlphaFold2 and Genetic Approaches","volume":"9","author":"Rodri\u0301guez Araya","year":"2023","journal-title":"ACS Infect. 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Cell. Proteomics"},{"issue":"12","key":"2026100909101416600_cit25","doi-asserted-by":"publisher","first-page":"1397","DOI":"10.15252\/embj.201386433","article-title":"Eaf5\/7\/3 Form\na Functionally Independent NuA4 Submodule Linked to RNA Polymerase\nII -coupled Nucleosome Recycling","volume":"33","author":"Rossetto","year":"2014","journal-title":"EMBO J."},{"issue":"9","key":"2026100909101416600_cit26","doi-asserted-by":"publisher","first-page":"2862","DOI":"10.1111\/cas.16263","article-title":"Understanding the Role of BRD8 in\nHuman Carcinogenesis","volume":"115","author":"Yamaguchi","year":"2024","journal-title":"Cancer Sci."},{"issue":"5","key":"2026100909101416600_cit27","doi-asserted-by":"publisher","first-page":"629","DOI":"10.1016\/j.chom.2016.04.020","article-title":"Epigenetic\nRegulation of Virulence Gene Expression in Parasitic Protozoa","volume":"19","author":"Duraisingh","year":"2016","journal-title":"Cell Host Microbe"},{"key":"2026100909101416600_cit28","doi-asserted-by":"publisher","first-page":"3963","DOI":"10.1093\/nar\/20.15.3963","article-title":"A Shuttle Vector Which\nFacilitates the Expression of Transfected Genes in Trypanosoma Cruzi\nand Leishmania","volume":"20","author":"Kelly","year":"1992","journal-title":"Nucleic Acids Res."},{"issue":"1","key":"2026100909101416600_cit29","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1042\/BJ20150986","article-title":"Glycosomal Bromodomain\nFactor 1 from Trypanosoma Cruzi Enhances Trypomastigote\nCell Infection\nand Intracellular Amastigote Growth","volume":"473","author":"Ritagliati","year":"2016","journal-title":"Biochem.\nJ."},{"key":"2026100909101416600_cit30","doi-asserted-by":"publisher","first-page":"5","DOI":"10.3389\/fcimb.2020.00005","article-title":"The Influence of Recombinational Processes to Induce\nDormancy in Trypanosoma Cruzi","volume":"10","author":"Resende","year":"2020","journal-title":"Front. 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Infect. 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Cell Sci."},{"issue":"11","key":"2026100909101416600_cit38","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0011731","article-title":"Morphogenesis\nin Trypanosoma Cruzi Epimastigotes Proceeds\nvia a Highly Asymmetric Cell Division","volume":"17","author":"Campbell","year":"2023","journal-title":"PLoS\nNeglected Trop. Dis."},{"issue":"4","key":"2026100909101416600_cit39","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1016\/j.mib.2010.05.006","article-title":"Assembly of\nthe Flagellum and Its Role in Cell Morphogenesis in Trypanosoma Brucei","volume":"13","author":"Vaughan","year":"2010","journal-title":"Curr. Opin. Microbiol."},{"issue":"23","key":"2026100909101416600_cit40","doi-asserted-by":"publisher","first-page":"24757","DOI":"10.1074\/jbc.M401276200","article-title":"The Trypanosoma\nBrucei Cyclin, CYC2,\nIs Required for Cell Cycle Progression through G1 Phase and for Maintenance\nof Procyclic Form Cell Morphology","volume":"279","author":"Hammarton","year":"2004","journal-title":"J. Biol.\nChem."},{"issue":"13","key":"2026100909101416600_cit41","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkaf661","article-title":"Comparative Mitochondrial Genome and Transcriptome Analyses Reveal\nStrain-Specific Features of RNA Editing in Trypanosoma Brucei","volume":"53","author":"Zhao","year":"2025","journal-title":"Nucleic Acids Res."},{"key":"2026100909101416600_cit42","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.34039","article-title":"Spontaneous\nDormancy Protects Trypanosoma Cruzi during Extended Drug Exposure","volume":"7","author":"Sa\u0301nchez-Valde\u0301z","year":"2018","journal-title":"eLife"},{"issue":"10","key":"2026100909101416600_cit43","doi-asserted-by":"publisher","first-page":"607","DOI":"10.1038\/s41579-019-0238-x","article-title":"Protozoan\nPersister-like Cells and Drug Treatment Failure","volume":"17","author":"Barrett","year":"2019","journal-title":"Nat. Rev. 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Microbiol."},{"key":"2026100909101416600_cit47","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2025.105413","article-title":"Trypanosoma Cruzi\nCell Cycle Progression Exhibits Minimal Variation in Histone PTMs\nwith Unique Histone H4 Acetylation Pattern","volume":"315","author":"Menezes","year":"2025","journal-title":"J. Proteomics"},{"issue":"5","key":"2026100909101416600_cit48","doi-asserted-by":"publisher","DOI":"10.1590\/0074-02760170404","article-title":"Trypanosoma\nCruzi Transcriptome during Axenic Epimastigote Growth Curve","volume":"113","author":"Santos","year":"2018","journal-title":"Mem. Inst. Oswaldo Cruz"},{"issue":"5","key":"2026100909101416600_cit49","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.00366-21","article-title":"Extensive Translational Regulation\nthrough the Proliferative Transition of Trypanosoma Cruzi Revealed\nby Multi-Omics","volume":"6","author":"Cha\u0301vez","year":"2021","journal-title":"mSphere"},{"issue":"11","key":"2026100909101416600_cit50","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0188441","article-title":"Transcriptome-Wide Analysis of the\nTrypanosoma Cruzi\nProliferative Cycle Identifies the Periodically Expressed mRNAs and\nTheir Multiple Levels of Control","volume":"12","author":"Cha\u0301vez","year":"2017","journal-title":"PLoS One"},{"issue":"1","key":"2026100909101416600_cit51","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.molbiopara.2015.11.001","article-title":"Expression of Non-Acetylatable Lysines\n10 and 14 of\nHistone H4 Impairs Transcription and Replication in Trypanosoma Cruzi","volume":"204","author":"Ramos","year":"2015","journal-title":"Mol. 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