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Chromatin structure: intro

Based on materials by Irina Zhegalova, Skoltech

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Levels of organization in space

Yadav et. al, 2018. Science

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C-based methods

Dekker et al., 2002. Science

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Chromatin matrix

Imakaev et al., 2012. Nature Methods; Lieberman-Aiden et al., 2009. Science

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3D genome

gure by S. Ulianov

TADs - Topologically Associated Domains

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3D genome

Structure

Compartments

0.5-10 Mb, Hi-C

0.1-1 Mb, Hi-C, 5C

50-500 kb, Hi-C, 4C, Capture-C

0.007-0.2 mkm2, Oligopaint-FISH

100-1000 nm2, ICH, Chrom-EMT

TADs

Loops

Nanodomains

Clutches

Putative functions

Editing strategy

Spatial segregation of active and inactive chromatin

Spatial insulation of neighboring genome loci and their regulatory systems

Spatial insulation of neighboring genome loci and contacts between regulatory regions

Unknown

Unknown

Unknown

Unknown

Unknown

Mutations and rearrangements of CBSs

Same as for TADs + CTCF recruitment

Size and assay

Identified by C-methods

Identified by microscopy

gure by S. Ulianov

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Trans-contacts

Bonev et al., 2016 Nature Reviews Genetics ; Hoencamp, 2021. Science

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Chromosome territories

Hi-C maps for chr 1, 2, 3 demonstrating territoriality:

  • Average number of contacts between pairs of chromosomes.
  • Average cis contacts are much higher than trans contacts.
  • cis-trans ratio as a QC step

Falk et al., 2019. Nature

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Compartments

Active promoters: H3K4me3, H3K9Ac

Active enhancers: H3K27Ac, H3K4me1

Repressors: H3K9me3, H3K27me3

Transcribed gene bodies: H3K36me3

Marchal, et al., 2019. Nature Reviews Molecular Cell Biology; gure by S. Ulianov

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Phase separation

Feric et al., 2021. Trends in Cell Biology

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Extrusion

SMC1

SMC3

RAD21

SA1

or SA2

Cohesin

CTCF

CTCF sites of the

same orientation

CTCF sites of

divergent orientation

CTCF sites of

convergent orientation

Extrusion

Extrusion

Zhang et al., 2022. NatRevImmunology ; Li et al., 2020. Nature

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TAD + TAD = compartment?

Starring:

cohesin as an Extruder CTCF as a Barrier

Knockout of CTCF results in TADs lost but not compartments

Compartments are less conserved across cell types than TADs

Phenotypic changes are more frequently associated with compartment repositioning than with structural disruption of loops or TADs

gure by S. Ulianov

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Loops

Dierent names for the same feature: loops, dots, enriched contacts

Rao et al., 2014. Cell gure by S. Ulyanov

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If there is no CTCF

Kristian Jeppsson, 2022 Science Advances; Heger et al., 2012. PNAS

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TADs vs loops

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What do they have in common?

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Chromatin Zoo

© 2023 I. Zhegalova OMICS

Dekker, J. et al., 2015. FEBS letters; Hoencamp, 2021. Science

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Chromatin structure: methods besides Hi-C

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Micro-C nucleosome scale

Hsieh et al., 2020. Molecular cell

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Single cell Hi-C

Flyamer et al., 2017. Nature

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Borders are stable

Ulianov et al., 2021. Nature Communications

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Red-C

RedChIP Red-C RADICL-seq

RNA-DNA SPRITE

Gavrilov et al., 2020. Nucleic Acids Research

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RNA-DNA SPRITE

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RNA-DNA SPRITE

Quinodoz et al., 2021. Cell

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Levels of organization in space

Yadav et. al, 2018. Science

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... and time

Cuvier and Fierz, 2017. Nature Reviews Genetics

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Microscopy - PALM

Xie et al., 2020. Nature Methods

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Oligopaint

optical reconstruction of chromatin architecture (ORCA)

method that uses

Oligopaint probes to label the genome with resolution reaching 2 kb

Mateo et al., 2021. Nature Protocols

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ORCA

Mateo et al., 2021. Nature Protocols

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HiGlass