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Real-Time Imaging of Translation on Single mRNA Transcripts in Live Cells

机译:实时成像在活细胞中单个mRNA转录物上的翻译

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摘要

Translation is under tight spatial and temporal controls to ensure protein production in the right time and place in cells. Methods that allow real-time, high-resolution visualization of translation in live cells are essential for understanding the spatiotemporal dynamics of translation regulation. Based on multivalent fluorescence amplification of the nascent polypeptide signal, we develop a method to image translation on individual mRNA molecules in real time in live cells, allowing direct visualization of translation events at the translation sites. Using this approach, we monitor transient changes of translation dynamics in responses to environmental stresses, capture distinct mobilities of individual polysomes in different subcellular compartments, and detect 3' UTR-dependent local translation and active transport of polysomes in dendrites of primary neurons.
机译:翻译受到严格的时空控制,以确保在正确的时间和位置在细胞中产生蛋白质。允许实时,高分辨率可视化翻译活细胞中的方法对于理解翻译调控的时空动态至关重要。基于新生多肽信号的多价荧光扩增,我们开发了一种在活细胞中实时对单个mRNA分子进行图像翻译的方法,可直接在翻译位点观察翻译事件。使用这种方法,我们监测翻译动力学对环境压力的响应的瞬时变化,捕获不同亚细胞区室中单个多核糖体的不同迁移率,并检测3'UTR依赖性局部翻译和多核糖体在初级神经元树突中的主动转运。

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