Abstract
Among the most important histone methyltransferases for metazoan development are EZH1/2 and their homologs, which methylate histone H3 lysine 27 and act as part of a highly conserved set of chromatin regulators called Polycomb Group (PcG) proteins. Reaching a precise understanding of the roles of PcG proteins in the orchestration of differentiation and the maintenance of cell identity requires a variety of genetic and molecular approaches. Here, we present a full suite of methods for the study of PcG proteins in early murine development, including mutant strain generation, embryonic stem cell derivation, epigenomic profiling, and immunofluorescence and in situ hybridization.
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Acknowledgments
We would like to thank Dr. Hiroki Sugishita for discussion and advice on CUT&Tag modifications. This work was supported by the Japan Agency for Medical Research and Development (AMED-CREST) (13417643 to H.K.), Grant-in-Aid for Scientific Research on Innovative Areas (JP19H05745 to H.K.), JSPS KAKENHI (JP19K22695, JP21K09784 to N.Y.-K.), and Takeda Science Foundation (to N.Y.-K.).
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Yakushiji-Kaminatsui, N., Kondo, T., Ohinata, Y., Takano, J., Koseki, H. (2022). Genetic, Genomic, and Imaging Approaches to Dissect the Role of Polycomb Group Epigenetic Regulators in Mice. In: Margueron, R., Holoch, D. (eds) Histone Methyltransferases. Methods in Molecular Biology, vol 2529. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2481-4_10
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DOI: https://doi.org/10.1007/978-1-0716-2481-4_10
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