Superresolution-Compatible DNA Labeling Technique with Silicon Rhodamine -Linked Nucleotide Reveals Chromatin Mobility and Organization Changes During Neuronal Differentiation
Datum
2025-01-10Beschreibung
Chromatin dynamics play crucial roles in cellular differentiation, yet tools for studying global chromatin mobility in living cells remain limited. Here, we developed a new STED-compatible silicon rhodamine-linked SiR-dCTP nucleotide combined with SNTT1 to label chromatin in live cells and track chromatin mobility during neural differentiation. Using correlative microscopy, we quantified the labeled chromatin domain sizes using STED super resolution and confocal and we have labeled domains of the sizes closer to chromatin loop domains. Additionally, using this approach we demonstrate that chromatin mobility progressively decreases during the transition from human induced pluripotent stem cells (iPSCs) to neural stem cells (NSCs) and ultimately to neurons. This reduction in mobility correlates with differentiation state, suggesting a potential role for chromatin dynamics in cellular plasticity. Mechanistic insights into this phenomenon using global MNase assay reveals that the accessibility of chromatin decreases during neuronal differentiation suggesting that chromatin gets more restricted with implications for understanding chromatin regulation during development.
DFG-Fächer
2.11-03 ZellbiologieZugehörige Drittmittelprojekte
DFG | CA198/20-1 | Regulation der SäugeDFG | SFB1361,TP06 | TP_06_Cardoso_Mainz
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