Abstract
In plants, chromatin dynamics spatiotemporally change in response to various environmental stimuli. However, little is known about chromatin dynamics in the nuclei of plants. Here, we introduce a three-dimensional, live-cell imaging method that can monitor chromatin dynamics in nuclei via a chromatin tagging system that can visualize specific genomic loci in living plant cells. The chromatin tagging system is based on a bacterial operator/repressor system in which the repressor is fused to fluorescent proteins. A recent refinement of promoters for the system solved the problem of gene silencing and abnormal pairing frequencies between operators. Using this system, we can detect the spatiotemporal dynamics of two homologous loci as two fluorescent signals within a nucleus and monitor the distance between homologous loci. These live-cell imaging methods will provide new insights into genome organization, development processes, and subnuclear responses to environmental stimuli in plants.
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Acknowledgements
This research was supported by CREST grants from the Japan Science and Technology Agency to S.M. and MEXT/JSPS KAKENHI to S.M. (No. 15H05962 and 26291067) and JSPS Research Fellowships to T.H. (No. 16J06389).
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Hirakawa, T., Matsunaga, S. (2016). Three-Dimensional, Live-Cell Imaging of Chromatin Dynamics in Plant Nuclei Using Chromatin Tagging Systems. In: Murata, M. (eds) Chromosome and Genomic Engineering in Plants. Methods in Molecular Biology, vol 1469. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4931-1_15
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DOI: https://doi.org/10.1007/978-1-4939-4931-1_15
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Publisher Name: Humana Press, New York, NY
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