Imaging nanoscale chromatin compaction in vivo

Non programmé
20m

Orateur

David LLERES (CNRS UMR5535)

Description

How metazoan genomes are structured at the nanoscale in living cells and tissues remains largely unknown. In addition, it still remains difficult to explore chromatin in vivo, particularly at both nucleosomal array level and single cell definition. Here, we applied a quantitative FRET (Forster resonance energy transfer)-based fluorescence lifetime imaging microscopy (FLIM) approach to measure nanoscale chromatin compaction in living primary cells and also at the scale of an organism, Caenorhabditis elegans. By measuring FRET between fluorescently-tagged core H2B histones, we spatially visualized distinct chromosomal regions and quantified the different levels of chromatin structuration.
In C.elegans, we combined RNAi approach to specifically define the heterochromatin state and showed that its architecture presents a nanoscale-compacted organization controlled by HP1 and SETDB1 H3-lysine-9 methyl-transferase homologs in vivo. Furthermore, we found that condensin I and condensin II regulate differentially the heterochromatin compaction state.
Altogether, our experimental system offers the exciting prospect to explore the effects of genetic and environmental factors on nanoscale chromatin compaction in living cells and in whole organisms.

Auteur principal

David LLERES (CNRS UMR5535)

Documents de présentation

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