Yunger Sharon, Shav-Tal Yaron
The Mina and Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan, Israel.
Methods Mol Biol. 2011;714:249-63. doi: 10.1007/978-1-61779-005-8_16.
The gene expression pathway begins in the nucleus as a gene receives a cue to transcribe, and typically ends in the cytoplasm with the production of the required protein. The nuclear processes of mRNA transcription and nucleo-cytoplasmic transport are of high importance as they encompass the major control points of gene expression. While it has been possible to study the mRNA life cycle using biochemical and molecular biology approaches, the advent of methods for nucleic acid tagging in vivo, have opened up many possibilities for examining these processes in vivo. In this chapter we describe the methodology required for setting up a live-cell system for monitoring real-time mRNA dynamics in mammalian cells.
基因表达途径始于细胞核,此时基因接收到转录信号,通常在细胞质中以所需蛋白质的产生而结束。mRNA转录和核质运输的核过程非常重要,因为它们包含了基因表达的主要控制点。虽然利用生化和分子生物学方法研究mRNA生命周期是可行的,但体内核酸标记方法的出现为在体内研究这些过程开辟了许多可能性。在本章中,我们描述了建立一个用于监测哺乳动物细胞中实时mRNA动态的活细胞系统所需的方法。