Profiling of H3K4me3 Modification in Plants using Cleavage under Targets and Tagmentation

作者： Xiaoyuan Tao*1,2, Mengtao Gao*3, Siyuan Wang*1, Xueying Guan1,4 1College of Agriculture and Biotechnology,Zhejiang University, 2Central Laboratory, State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products,Zhejiang Academy of Agricultural Sciences, 3State Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Hybrid R & D Engineering Center (the Ministry of Education), College of Agriculture,Nanjing Agricultural University, 4Hainan Institute of Zhejiang University

简介：

Overview

This study examines the refined application of the CUT&Tag method for epigenomic profiling in plants, focusing on histone modifications. The primary goal is to effectively isolate and analyze protein-affiliated DNA fragments within plant genomes.

Key Study Components

Research Area

• Chromatin epigenomics
• Plant biology
• Histone modifications

Background

• Identifying protein-affiliated DNA fragments to assess chromatin status
• Importance of histone modifications in gene expression regulation
• Development of CUT&Tag technology for efficient epigenomic profiling

Methods Used

• Isolation of intact nuclei from plant cells (cotton leaves)
• Use of specific antibodies for chromatin modifications
• Tn5 transposase for fragmenting chromatin in situ

Main Results

• Successful isolation of intact nuclei and profiling of histone H3 Lysine-4 Trimethylation
• Enhanced DNA preparation and sequencing through PCR enrichment
• Validated efficiency of the refined CUT&Tag protocol

Conclusions

• The study successfully demonstrates a refined CUT&Tag protocol for epigenomic profiling in plants.
• Findings contribute valuable insights into the molecular mechanisms of gene expression regulation.

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