Microscopic analysis of chromatin localization and dynamics in C. elegans

Lanctôt, Christian; Meister, Pierre (2013). Microscopic analysis of chromatin localization and dynamics in C. elegans. In: Shav-Tal, Yaron (ed.) Imaging Gene Expression. Methods in Molecular Biology: Vol. 1042 (pp. 153-172). New York: Humana Press 10.1007/978-1-62703-526-2_11

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During development, the genome undergoes drastic reorganization within the nuclear space. To determine tridimensional genome folding, genome-wide techniques (damID/Hi-C) can be applied using cell populations, but these have to be calibrated using microscopy and single-cell analysis of gene positioning. Moreover, the dynamic behavior of chromatin has to be assessed on living samples. Combining fast stereotypic development with easy genetics and microscopy, the nematode C. elegans has become a model of choice in recent years to study changes in nuclear organization during cell fate acquisition. Here we present two complementary techniques to evaluate nuclear positioning of genes either by fluorescence in situ hybridization in fixed samples or in living worm embryos using the GFP-lacI/lacO chromatin-tagging system.

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