通过显微注射评估哺乳动物细胞中的 mRNA 核输出。

Assessing mRNA nuclear export in mammalian cells by microinjection.

机构信息

Department of Biochemistry, University of Toronto, Toronto, ON, Canada.

出版信息

Methods. 2017 Aug 15;126:76-85. doi: 10.1016/j.ymeth.2017.05.027. Epub 2017 Jun 1.

DOI:10.1016/j.ymeth.2017.05.027

Abstract

The nuclear export of mRNAs is an important yet little understood part of eukaryotic gene expression. One of the easiest methods for monitoring mRNA export in mammalian tissue culture cells is through the microinjection of DNA plasmids into the nucleus and monitoring the distribution of the transcribed product over time. Here we describe how to setup a microscope equipped with a micromanipulator used in cell microinjections, and we explain how to perform a nuclear mRNA export assay and obtain the nuclear export rate for any given mRNA.

摘要

mRNA 的核输出是真核基因表达中一个重要但了解甚少的部分。在哺乳动物组织培养细胞中监测 mRNA 输出的最简单方法之一是将 DNA 质粒显微注射到细胞核中，并随时间监测转录产物的分布。这里我们描述了如何设置配备用于细胞显微注射的微操作器的显微镜，并解释了如何进行核 mRNA 输出测定并获得任何给定 mRNA 的核输出率。