Products/Services Used | Details | Operation |
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PCR Cloning and Subcloning> | Lentiviral vectors expressing C-terminally eGFP-tagged Cenpa and Trf1 under EF1a promoter were generated by Genscript custom gene service by subcloning Cenpa ORF (#OMu00596C, NM_007681.3) and Trf1 ORF (#OMu19848C, NM_009352.3) into pLENTI-dCas9-VP64_GFP backbone (Konermann et al., 2015). | Get A Quote |
Nucleosomes form heterogeneous groups in vivo, named clutches. Clutches are smaller and less dense in mouse embryonic stem cells (ESCs) compared to neural progenitor cells (NPCs). Using coarse-grained modeling of the pluripotency Pou5f1 gene, we show that the genome-wide clutch differences between ESCs and NPCs can be reproduced at a single gene locus. Larger clutch formation in NPCs is associated with changes in the compaction and internucleosome contact probability of the Pou5f1 fiber. Using single-molecule tracking (SMT), we further show that the core histone protein H2B is dynamic, and its local mobility relates to the structural features of the chromatin fiber. H2B is less stable and explores larger areas... More