Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Japan.
Methods Mol Biol. 2023;2577:103-122. doi: 10.1007/978-1-0716-2724-2_8.
The Spliced TetO REpeAt, MS2 repeat, and INtein sandwiched reporter Gene tag (STREAMING-tag) system enables imaging of nuclear localization as well as the transcription activity of a specific endogenous gene at sub-100-nm resolution in living cells. The use of this system combined with imaging of epigenome states enables a detailed analysis of the impact of epigenome status on transcriptional dynamics. In this chapter, we describe a method for quantifying distances between Nanog gene and clusters of cofactor BRD4 using the STREAMING-tag system in mouse embryonic stem cells.
拼接 TetO REpeAt、MS2 重复序列和内含肽夹合报告基因标签(STREAMING-tag)系统可在活细胞中以亚 100nm 分辨率对核定位以及特定内源性基因的转录活性进行成像。该系统与表观基因组状态成像相结合,可对表观基因组状态对转录动力学的影响进行详细分析。在本章中,我们描述了一种在小鼠胚胎干细胞中使用 STREAMING-tag 系统定量 Nanog 基因和共因子 BRD4 簇之间距离的方法。