RNA Detection and Visualization pp 221-235
In Vivo Visualization of RNA Using the U1A-Based Tagged RNA System
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.
Key wordsU1A GFP Tagged RNA RNA localization ASH1 RNP (ribonucleoprotein) complex
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