STREAMING-Tag System: Technology to Enable Visualization of Transcriptional Activity and Subnuclear Localization of Specific Endogenous Genes

Hiroaki Ohishi; Hiroshi Ochiai

doi:10.1007/978-1-0716-2724-2_8

STREAMING-Tag System: Technology to Enable Visualization of Transcriptional Activity and Subnuclear Localization of Specific Endogenous Genes

Methods Mol Biol. 2023:2577:103-122. doi: 10.1007/978-1-0716-2724-2_8.

Authors

Hiroaki Ohishi¹, Hiroshi Ochiai²

Affiliations

¹ Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Japan.
² Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Japan. ochiai@hiroshima-u.ac.jp.

PMID: 36173569
DOI: 10.1007/978-1-0716-2724-2_8

Abstract

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.

Keywords: Gene regulation; ImageJ; Live imaging; Python; RNAPII; STREAMING-tag; Trackpy; Transcriptional bursting; Transcriptional regulatory factor.

MeSH terms

Animals
Mice
Mouse Embryonic Stem Cells
Nuclear Proteins
Technology
Transcription Factors* / genetics
Transcription, Genetic*

Substances

Nuclear Proteins
Transcription Factors