In 2019, Kaya-Okur et al. reported on the cleavage under targets and tagmentation (CUT&Tag) technology for efficient profiling of epigenetically modified DNA fragments. It was used mainly for cultured cell lines and was especially effective for small samples and single cells. This strategy generated high-resolution and low-background-noise chromatin profiling data for epigenomic analysis. CUT&Tag is well suited to be used in plant cells, especially in tissues from which small samples are taken, such as ovules, anthers, and fibers. Here, we present a CUT&Tag protocol step by step using plant nuclei. In this protocol, we quantified the nuclei that can be used in each CUT&Tag reaction, and compared the efficiency of CUT&Tag with chromatin immunoprecipitation with sequencing (ChIP-seq) in the leaves of cotton. A general workflow for the bioinformatic analysis of CUT&Tag is also provided. Results indicated that, compared with ChIP-seq, the CUT&Tag procedure was faster and showed a higher-resolution, lower-background signal than did ChIP. A CUT&Tag protocol has been refined for plant cells using intact nuclei that have been isolated.

中文翻译：

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使用 CUT&Tag 对棉花中 H3K4me3 修饰进行有效染色质分析。

2019 年，Kaya-Okur 等人。报道了用于有效分析表观遗传修饰的 DNA 片段的靶点切割和标记 (CUT&Tag) 技术。它主要用于培养的细胞系，对小样本和单细胞特别有效。该策略为表观基因组分析生成了高分辨率和低背景噪声的染色质分析数据。CUT&Tag 非常适合用于植物细胞，尤其是用于提取小样本的组织，例如胚珠、花药和纤维。在这里，我们使用植物核逐步介绍了 CUT&Tag 协议。在该协议中，我们量化了可用于每个 CUT&Tag 反应的细胞核，并将 CUT&Tag 与染色质免疫沉淀和测序 (ChIP-seq) 在棉花叶子中的效率进行了比较。还提供了 CUT&Tag 生物信息学分析的一般工作流程。结果表明，与 ChIP-seq 相比，CUT&Tag 过程比 ChIP 更快，并且显示出更高的分辨率、更低的背景信号。使用已分离的完整细胞核针对植物细胞改进了 CUT&Tag 协议。