Chung Sunglan, Takizawa Peter A
Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA.
Methods Mol Biol. 2011;714:221-35. doi: 10.1007/978-1-61779-005-8_14.
mRNA transport is a widely used method to achieve the asymmetric distribution of proteins within a cell or organism. In order to understand how RNA is transported, it is essential to utilize a system that can readily detect RNA movement in live cells. The tagged RNA system has recently emerged as a feasible non-invasive solution for such purpose. In this chapter, we describe in detail the U1A-based tagged RNA system. This system coexpresses U1Ap-GFP with the RNA of interest tagged with U1A aptamers, and has been proven to effectively track RNA in vivo. In addition, we provide further applications of the system for ribonucleoprotein complex purification by TAP-tagging the U1Ap-GFP construct.
信使核糖核酸（mRNA）运输是一种广泛应用的方法，用于在细胞或生物体内实现蛋白质的不对称分布。为了理解RNA是如何运输的，利用一个能够在活细胞中轻松检测RNA移动的系统至关重要。标记RNA系统最近已成为用于此目的的一种可行的非侵入性解决方案。在本章中，我们详细描述基于U1A的标记RNA系统。该系统将U1Ap-绿色荧光蛋白（GFP）与用U1A适体标记的目标RNA共表达，并且已被证明能在体内有效追踪RNA。此外，我们还介绍了通过对U1Ap-GFP构建体进行串联亲和纯化（TAP）标记，该系统在核糖核蛋白复合物纯化方面的进一步应用。
