Information about the positioning of individual loci in the nucleus and the status of epigenetic modifications at these loci in each cell contained in plant tissue increases our understanding of how cells in a tissue coordinate gene expression. To obtain such information, a less damaging method of visualizing DNA in tissue that can be used with immunohistochemistry is required. Recently, a less damaging DNA visualization method using the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/associated caspase 9) system, named RNA-guided endonuclease–in situ labeling (RGEN-ISL), was reported. This system made it possible to visualize a target DNA locus in a nucleus fixed on a glass slide with a set of simple operations, but it could not be applied to cells in plant tissues. In this work, we have developed a modified RGEN-ISL method with decrosslinking that made it possible to simultaneously detect the DNA loci and immunohistochemistry signals, including histone modification, in various types of plant tissues and species.

中文翻译：

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解交联可以实现植物组织中 RNA 引导核酸内切酶原位标记信号的可视化


有关细胞核中各个基因座的定位以及植物组织中每个细胞中这些基因座的表观遗传修饰状态的信息，增加了我们对组织中细胞如何协调基因表达的理解。为了获得此类信息，需要一种可与免疫组织化学一起使用的破坏性较小的组织中 DNA 可视化方法。最近，报道了一种使用 CRISPR/Cas9（成簇规则间隔短回文重复序列/相关 caspase 9）系统的破坏性较小的 DNA 可视化方法，称为 RNA 引导核酸内切酶原位标记 (RGEN-ISL)。该系统可以通过一系列简单的操作来可视化固定在载玻片上的细胞核中的目标DNA位点，但它不能应用于植物组织中的细胞。在这项工作中，我们开发了一种改良的 RGEN-ISL 解交联方法，可以同时检测各种类型的植物组织和物种中的 DNA 位点和免疫组织化学信号，包括组蛋白修饰。