Three dimensional, live-cell imaging of chromosome dynamics in plant nuclei using chromatin tagging systems.
Three dimensional, live-cell imaging of chromosome dynamics in plant nuclei using chromatin tagging systems.
复制标题
使用染色质标记系统对植物细胞核中染色体动力学进行三维活细胞成像。
DOI：
10.1007/978-1-4939-4931-1_15
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发表时间：
2016
影响因子：
--
通讯作者：
S.
中科院分区：
文献类型：
--
作者：
Hirakawa;T. and Matsunaga;S.
在植物中，染色质动力学响应于各种环境刺激而发生时空变化。然而，对植物细胞核中的染色质动力学知之甚少。在这里，我们介绍了一种三维活细胞成像方法，可以通过染色质标记系统监测细胞核中的染色质动力学，该系统可以可视化活植物细胞中的特定基因组位点。染色质标记系统是基于细菌操作符/阻遏物系统，其中阻遏物融合到荧光蛋白。最近对系统启动子的改进解决了基因沉默和操作符之间异常配对频率的问题。利用该系统，我们可以检测两个同源基因座在细胞核内作为两个荧光信号的时空动态，并监测同源基因座之间的距离。这些活细胞成像方法将为植物基因组组织、发育过程和对环境刺激的亚核反应提供新的见解。
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.