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Super-resolution imaging of chromatin and nuclear organisation (Rob White, Steven Lee)

Supervisors: Rob White, Steven Lee

The 3-D organization of chromatin within the nucleus is likely to play a major role in defining cell fates with the specific sub-nuclear localisation of active and inactive genes. The new techniques of super-resolution microscopy offer powerful approaches to investigate 3-D chromatin organization. In this project we will apply super-resolution microscopy to investigate the organization of the Drosophila spermatocyte nucleus. These nuclei have key advantages for imaging as they are up to 25X the volume of normal somatic cell nuclei and individual chromosomes and chromatin fibers are easily visualized. In addition they are undergoing a clear well-characterised transcriptional program with the specific expression of several thousand spermatogenesis genes and we have many reagents available to probe 3-D chromatin organization in these nuclei. We will focus on visualization of the domain organization of chromatin and its association with chromatin states. We will use genetic approaches to study the regulation of 3D chromatin organization and tagged-loci to study the dynamics of gene localization in the nucleus and its association with the regulation of gene expression.

Relevant references

Super-resolution imaging of the nucleoid-associated protein HU in Caulobacter crescentus (2011) SF Lee, MA Thompson, MA Schwartz, L Shapiro, WE Moerner – Biophys J 100, L31.

Redhouse, J. L., Mozziconacci, J. and White, R. (2011) Co-transcriptional architecture in a Y loop in Drosophila melanogaster. Chromosoma 12: 0399-407.

White, R. (2012) Packaging the fly genome: domains and dynamics. Briefings in Functional Genomics 11:347-55.

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