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Table of contents
Introduction
1 On Chromatin Architecture
1.1 Chromatin structure and function
1.1.1 Chromatin structure: how to characterize it
1.1.2 Chromatin structures span over different length-scales
1.2 On Topologically Associating Domains (TADs)
1.2.1 How are TAD formed: the loop extrusion hypothesis
1.3 CTCF and Cohesin: a focus on chromatin organisers
1.3.1 CTCF
1.3.2 Cohesin
1.3.3 CTCF, Cohesin and chromatin structure
1.4 Goal of this work
2 Technique and system
2.1 Optical microscopy
2.1.1 Diffraction limit and fluorescence
2.1.2 Single Molecule tracking
2.2 Microscopy set-up
2.3 Biological system
2.3.1 Cell Culture
2.4 The experiments
2.4.1 The degron system
2.4.2 Labelling and imaging conditions
2.5 Analysis of single molecule imaging
2.5.1 Analysis of binding kinetics
2.5.2 Residence time
2.5.3 Analysis of dynamics
3 Results and discussion
3.1 Chromatin as a reference
3.2 CTCF
3.3 Cohesin
3.4 Cohesin without CTCF
3.4.1 Cohesin in non-cycling cells
3.5 Other mutants
3.5.1 Cohesin in absence of Sororin
3.5.2 Cohesin in absence of Nipbl (Scc2)
3.5.3 Control
3.6 Discussion
3.6.1 CTCF
3.6.2 Cohesin
3.7 Conclusion and perspectives
Acknowledgements
References
