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© Research
Publication : Genome biology

Super-resolution visualization and modeling of human chromosomal regions reveals cohesin-dependent loop structures.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Genome biology - 11 May 2021

Hao X, Parmar JJ, Lelandais B, Aristov A, Ouyang W, Weber C, Zimmer C,

Link to Pubmed [PMID] – 33975635

Link to DOI – 10.1186/s13059-021-02343-w

Genome Biol 2021 May; 22(1): 150

The 3D organization of the chromatin fiber in cell nuclei plays a key role in the regulation of gene expression. Genome-wide techniques to score DNA-DNA contacts, such as Hi-C, reveal the partitioning of chromosomes into epigenetically defined active and repressed compartments and smaller “topologically associated” domains. These domains are often associated with chromatin loops, which largely disappear upon removal of cohesin. Because most Hi-C implementations average contact frequencies over millions of cells and do not provide direct spatial information, it remains unclear whether and how frequently chromatin domains and loops exist in single cells.We combine 3D single-molecule localization microscopy with a low-cost fluorescence labeling strategy that does not denature the DNA, to visualize large portions of single human chromosomes in situ at high resolution. In parallel, we develop multi-scale, whole nucleus polymer simulations, that predict chromatin structures at scales ranging from 5 kb up to entire chromosomes. We image chromosomes in G1 and M phase and examine the effect of cohesin on interphase chromatin structure. Depletion of cohesin leads to increased prevalence of loose chromatin stretches, increased gyration radii, and reduced smoothness of imaged chromatin regions. By comparison to model predictions, we estimate that 6-25 or more purely cohesin-dependent chromatin loops coexist per megabase of DNA in single cells, suggesting that the vast majority of the genome is enclosed in loops.Our results provide new constraints on chromatin structure and showcase an affordable non-invasive approach to study genome organization in single cells.

https://pubmed.ncbi.nlm.nih.gov/33975635