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The nucleus in 4D

Organizer:

Ortrun Mittelsten Scheid

Background:

Genetic maps or genome browsers illustrate chromosomes as linear units, but this is of course far from reality. The long chromosomal DNA molecules undergo condensation in the order of several magnitudes, while keeping flexibility and accessibility for the numerous DNA-related processes. The arrangement of the genome within the nucleus is not random in most organisms, but individual regions can change their location over long distances. Structural aspects of eukaryotic nuclei have been investigated since the early days of light microscopy, but research on functional aspects of spatial nuclear organization is a relatively new field. Complementary sophisticated technologies have recently allowed exciting insights into nuclear substructures down to the molecular level, and we consider this topic indispensable in the frame of this DK.

Objectives:

To familiarize DK students with different approaches, we plan

  • To present the principles and examples of chromosome conformation capture techniques
  • To present techniques to analyse interaction of genome regions with the nuclear envelope
  • To introduce state-of-the-art cytological techniques
  • To make the students discuss the potential and the limits of the approaches
  • To discuss several models of nuclear organization across a range of organisms

 

Contents:
  • Basic principles of chromosome conformation capture (3C, 4C, Hi-C)
  • Basic principles of DNA footprinting techniques to reveal interactions with nuclear structures including envelope and pores (DamID and related methods)
  • Basic principles of nuclear structure analysis (chromosome territories, 3D imaging techniques by high resolution microscopy of nuclei from fixed and living cells)
  • Basic principles of tracking spatial and temporal location of individual molecules or regions (RNA/DNA FISH, beacon tracking, visualization by CRISPR/Cas fusions)
  • Students prepare to “advocate” selected techniques and to challenge the experts with questions and arguments
  • Experts share their vision and discuss potential future directions of this research field with the students

 

MFPL

 

 

UniVie

 

 

GMI

 

 

IMBA Logo

 

 

FWF