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| Content Provider | Springer Nature : BioMed Central |
|---|---|
| Author | Hao, Xian Parmar, Jyotsana J. Lelandais, Benoît Aristov, Andrey Ouyang, Wei Weber, Christian Zimmer, Christophe |
| Abstract | Background 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. Results 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. Conclusion Our results provide new constraints on chromatin structure and showcase an affordable non-invasive approach to study genome organization in single cells. |
| Related Links | https://genomebiology.biomedcentral.com/counter/pdf/10.1186/s13059-021-02343-w.pdf |
| Ending Page | 20 |
| Page Count | 20 |
| Starting Page | 1 |
| File Format | HTM / HTML |
| DOI | 10.1186/s13059-021-02343-w |
| Journal | Genome Biology |
| Issue Number | 1 |
| Volume Number | 22 |
| Language | English |
| Publisher | BioMed Central |
| Publisher Date | 2021-05-11 |
| Access Restriction | Open |
| Subject Keyword | Animal Genetics and Genomics Human Genetics Plant Genetics and Genomics Microbial Genetics and Genomics Bioinformatics Evolutionary Biology Chromatin Chromosomes Cohesin Super-resolution microscopy Polymer models |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biotechnology Genetics |
| Journal Impact Factor | 10.1/2023 |
| 5-Year Journal Impact Factor | 16.5/2023 |
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