biostudies-literatureGuo D"Xingdian Talent Support Program" of Yunnan ProvinceYunnan Fundamental Research ProjectsNatural Science Foundation of Yunnan ProvinceNational Natural Science Foundation of China“Xingdian Talent Support Program” of Yunnan Provincegkaf302https://www.ebi.ac.uk/biostudies/studies/S-EPMC11995263biostudies-literatureUnknown53(7)Two-cell-like cells (2CLCs), a rare population (∼0.5%) in mouse embryonic stem cell (mESC) cultures, are in a transient totipotent-like state resembling that of 2C-stage embryos, and their discovery and characterization have greatly facilitated the study of early developmental events, such as zygotic genome activation. However, the molecular determinants governing 2C-like reprogramming remain to be elucidated. Here, we show that ZBTB24, CDCA7, and HELLS, components of a molecular pathway that is involved in the pathogenesis of immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome, function as negative regulators of 2C-like reprogramming by maintaining DNA methylation of the Dux cluster, a master inducer of the 2C-like state. Disruption of the ZBTB24-CDCA7-HELLS axiNucleic acids researchThe ZBTB24-CDCA7-HELLS axis suppresses the totipotent 2C-like reprogramming by maintaining Dux methylation and repression.PMC11995263202301AT070304202201BE070001-006KKRD202273103202102AA10005332270845Dan JLiu YZhang JLu YHardikar SGuo DDu ZLin MXue YChen TfalseThe ZBTB24-CDCA7-HELLS axis suppresses the totipotent 2C-like reprogramming by maintaining Dux methylation and repression.Two-cell-like cells (2CLCs), a rare population (∼0.5%) in mouse embryonic stem cell (mESC) cultures, are in a transient totipotent-like state resembling that of 2C-stage embryos, and their discovery and characterization have greatly facilitated the study of early developmental events, such as zygotic genome activation. However, the molecular determinants governing 2C-like reprogramming remain to be elucidated. Here, we show that ZBTB24, CDCA7, and HELLS, components of a molecular pathway that is involved in the pathogenesis of immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome, function as negative regulators of 2C-like reprogramming by maintaining DNA methylation of the Dux cluster, a master inducer of the 2C-like state. Disruption of the ZBTB24-CDCA7-HELLS axi2025-01-01T00:00:00Z2025 Apr2026-07-15T08:56:09.919Z2026-07-02T03:08:38.854ZS-EPMC119952634022691810.1093/nar/gkaf302