Fujiwara Yasuhiro, Okada Yuki
Institute for Quantitative Biosciences, The University of Tokyo, Tokyo, Japan.
Methods Mol Biol. 2023;2519:141-153. doi: 10.1007/978-1-0716-2433-3_16.
Epigenome research has employed various methods to identify the genomic location of proteins of interest, such as transcription factors and histone modifications. CUT&Tag is a recently established method used in epigenome research to determine the genomic location of proteins of interest, such as transcription factors and histone modifications. In CUT&Tag method, cells are bound and hold on concanavalin A (con A)-coated magnetic beads, then a Protein-A Tn5 transposase fusion protein cuts the genome and inserts adapter sequences nearby the target protein. Here we describe the updated CUT&Tag procedure using "home-made" con A-conjugated magnetic beads. This method is free of poor suspendability and severe aggregation, hence providing improved sensitivity.
表观基因组研究采用了多种方法来确定感兴趣的蛋白质（如转录因子和组蛋白修饰）的基因组位置。CUT&Tag是表观基因组研究中最近建立的一种用于确定感兴趣蛋白质（如转录因子和组蛋白修饰）基因组位置的方法。在CUT&Tag方法中，细胞与伴刀豆球蛋白A（Con A）包被的磁珠结合并固定，然后一种蛋白A-Tn5转座酶融合蛋白切割基因组并在靶蛋白附近插入接头序列。在此，我们描述了使用“自制”Con A偶联磁珠的更新后的CUT&Tag程序。该方法没有悬浮性差和严重聚集的问题，因此提高了灵敏度。
