Tao Xiaoyuan, Feng Shouli, Zhao Ting, Guan Xueying
College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, 310058 China.
Plant Methods. 2020 Aug 31;16:120. doi: 10.1186/s13007-020-00664-8. eCollection 2020.
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.
2019年，卡亚-奥库尔等人报道了靶向切割与片段化标记（CUT&Tag）技术，用于对表观遗传修饰的DNA片段进行高效分析。该技术主要用于培养的细胞系，对小样本和单细胞尤为有效。此策略为表观基因组分析生成了高分辨率、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，特别是从小样本取材的组织，如胚珠、花药和纤维。
在此，我们逐步展示了一种使用植物细胞核的CUT&Tag实验方案。在该方案中，我们对每个CUT&Tag反应中可使用的细胞核进行了定量，并在棉花叶片中比较了CUT&Tag与染色质免疫沉淀测序（ChIP-seq）的效率。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操作更快，且显示出比ChIP更高的分辨率、更低的背景信号。
已针对植物细胞优化了一种使用分离出的完整细胞核的CUT&Tag实验方案。
