Abstract
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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Acknowledgements
We thank Darina Korčeková for expert help in developing 3D DNA FISH protocols, the Meister laboratory, Susan Gasser, and the Gasser laboratory for continuous support and helpful discussions. This work was funded in part by programs of the Charles University in Prague (UNCE 204022 and Prvouk/1LF/1) as well as by the Czech Science Foundation (grants P302/11/1262 and P302/12/G157), the Swiss National Foundation (SNF assistant professor grant PP00P3_133744), and the Fondation Suisse pour le Recherche sur les Maladies Musculaires.
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Lanctôt, C., Meister, P. (2013). Microscopic Analysis of Chromatin Localization and Dynamics in C. elegans . In: Shav-Tal, Y. (eds) Imaging Gene Expression. Methods in Molecular Biology, vol 1042. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-526-2_11
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DOI: https://doi.org/10.1007/978-1-62703-526-2_11
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