Profiling of H3K4me3 Modification in Plants using Cleavage under Targets and Tagmentation
文献类型： 外文期刊
作者： Tao, Xiaoyuan 1 ; Gao, Mengtao 3 ; Wang, Siyuan 1 ; Guan, Xueying 1 ;
作者机构： 1.Zhejiang Univ, Coll Agr & Biotechnol, Hangzhou, Zhejiang, Peoples R China
2.Zhejiang Acad Agr Sci, Cent Lab, State Key Lab Managing Biot & Chem Threats Qual &, Hangzhou, Zhejiang, Peoples R China
3.Nanjing Agr Univ, Coll Agr, Minist Educ,Cotton Hybrid R&D Engn Ctr, State Key Lab Crop Genet & Germplasm Enhancement, Nanjing, Peoples R China
4.Zhejiang Univ, Hainan Inst, Hangzhou, Zhejiang, Peoples R China
期刊名称：JOVE-JOURNAL OF VISUALIZED EXPERIMENTS （ 影响因子：1.424； 五年影响因子：1.789 ）
ISSN： 1940-087X
年卷期： 2022 年 182 期
页码：
收录情况： SCI
摘要： Epigenomic regulation at the chromatic level, including DNA and histone modifications, behaviors of transcription factors, and non-coding RNAs with their recruited proteins, lead to temporal and spatial control of gene expression. Cleavage under targets and tagmentation (CUT&Tag) is an enzyme-tethering method in which the specific chromatin protein is firstly recognized by its specific antibody, and then the antibody tethers a protein A-transposase (pA-Tn5) fusion protein, which cleaves the targeted chromatin in situ by the activation of magnesium ions. Here, we provide our previously published CUT&Tag protocol using intact nuclei isolated from allortetraploid cotton leaves with modification. This step-by-step protocol can be used for epigenomic research in plants. In addition, substantial modifications for plant nuclei isolation are provided with critical comments.
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