用于特定内源性基因转录活性和核定位实时成像的定位与表达实时观察（ROLEX）系统

Real-Time Observation of Localization and Expression (ROLEX) System for Live Imaging of the Transcriptional Activity and Nuclear Position of a Specific Endogenous Gene.

作者信息

Ochiai Hiroshi

机构信息

JST, PRESTO, Higashi-Hiroshima, Japan.

Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Japan.

出版信息

Methods Mol Biol. 2019;2038:35-45. doi: 10.1007/978-1-4939-9674-2_3.

DOI:10.1007/978-1-4939-9674-2_3

PMID:31407276

Abstract

Long genomic DNA is folded in a cell-type-specific manner and stored in the cell nucleus. The higher-order structure of genomic DNA is thought to be important for DNA transcription, repair, and replication. Recent advancements in live cell imaging techniques that enable the labeling of specific genomic loci and RNA have made it possible to capture the dynamic relationships between higher-order genomic structure and gene expression. We have established the real-time observation of localization and expression (ROLEX) system for live imaging of the transcriptional state and nuclear position of a specific endogenous gene. In this chapter, I will introduce the detailed protocol of ROLEX imaging in mouse embryonic stem cells.

摘要

长链基因组DNA以细胞类型特异性方式折叠并存储在细胞核中。基因组DNA的高级结构被认为对DNA转录、修复和复制很重要。能够标记特定基因组位点和RNA的活细胞成像技术的最新进展使得捕捉高级基因组结构与基因表达之间的动态关系成为可能。我们已经建立了用于实时观察特定内源基因转录状态和核位置的定位与表达实时观察（ROLEX）系统。在本章中，我将介绍在小鼠胚胎干细胞中进行ROLEX成像的详细方案。