Superresolution-Compatible DNA Labeling Technique with Silicon Rhodamine -Linked Nucleotide Reveals Chromatin Mobility and Organization Changes During Neuronal Differentiation

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dc.contributor.author Pabba, Maruthi K.
dc.contributor.author Kraus, Tomáš
dc.contributor.author Meyer, Janis
dc.contributor.author Celikay, Kerem
dc.contributor.author Maiser, Andreas
dc.contributor.author Leonhardt, Heinrich
dc.contributor.author Rohr, Karl
dc.contributor.author Hocek, Michal
dc.contributor.author Cardoso, M. Cristina
dc.contributor.author Harz, Hartmann
dc.contributor.author Pradhan, Sunil Kumar
dc.contributor.author Kuba, Miroslav
dc.date.accessioned 2025-01-23T17:54:48Z
dc.date.available 2025-01-23T17:54:48Z
dc.date.created 2025-01-10
dc.date.issued 2025-01-23
dc.description Chromatin dynamics play a crucial role in cellular differentiation, yet tools for studying global chromatin mobility in living cells remain limited. Here, we developed a novel probe for the metabolic labeling of chromatin and tracking its mobility during neural differentiation. The labeling system utilizes a newly developed silicon rhodamine-conjugated deoxycytidine triphosphate (dCSiRTP). We show that this dCTP is efficiently delivered into living human induced pluripotent stem cells (iPSCs) and neural stem cells (NSCs) via a synthetic transporter (SNTT1). Using correlative confocal microscopy and stimulated emission depletion (STED) super-resolution microscopy, we quantified the sizes of labeled chromatin domains. Time-lapse super-resolution microscopy combined with single particle tracking revealed that chromatin mobility decreases during the transition from iPSCs (pluripotent state) to NSCs and neurons (differentiated state). This reduction in mobility correlates with the differentiation state, reflecting changes in chromatin organization during cell fate commitment. Concomitant mechanistic insights obtained from micrococcal nuclease digestion assays, chromatin compaction and histone modification analyses revealed a decrease in chromatin accessibility during neuronal differentiation. These data indicate that chromatin adopts a more constrained structure with reduced accessibility and increased heterochromatin-associated histone modifications. These findings provide new insights into chromatin regulation during neurogenesis. de_DE
dc.identifier.uri https://tudatalib.ulb.tu-darmstadt.de/handle/tudatalib/4441
dc.identifier.uri https://doi.org/10.48328/tudatalib-1651
dc.language.iso en de_DE
dc.rights.licenseCC-BY-NC-4.0 (https://creativecommons.org/licenses/by-nc/4.0)
dc.subject SIR-dCTP de_DE
dc.subject SNTT1 de_DE
dc.subject chromatin mobility de_DE
dc.subject neural differentiation de_DE
dc.subject STED de_DE
dc.subject.classification 2.11-03
dc.subject.ddc 570
dc.title Superresolution-Compatible DNA Labeling Technique with Silicon Rhodamine -Linked Nucleotide Reveals Chromatin Mobility and Organization Changes During Neuronal Differentiation de_DE
dc.type Text de_DE
dc.type Software de_DE
dc.type Image de_DE
dcterms.accessRights openAccess
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tuda.project DFG | CA198/20-1 | Regulation der Säuge
tuda.project DFG | SFB1361,TP06 | TP_06_Cardoso_Mainz
tuda.unit TUDa

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