Li Ying, Aggarwal Mahima B, Ke Ke, Nguyen Kim, Spitale Robert C
Biochemistry. 2018 Mar 13;57(10):1577-1581. doi: 10.1021/acs.biochem.8b00053. Epub 2018 Feb 27.
Recent analysis of transcriptomes has revealed that RNAs perform a myriad of functions beyond encoding proteins. Critical to RNA function is its transport to unique subcellular locations. Despite the importance of RNA localization, it is still very challenging to study in an unbiased manner. We recently described the ability to tag RNA molecules within subcellular locations through spatially restricted nucleobase oxidation. Herein, we describe a dramatic improvement of this protocol through the localized oxidation and tagging of proteins. Isolation of RNA-protein complexes enabled the enrichment of challenging RNA targets on chromatin and presented a considerably optimized protocol for the analysis of RNA subcellular localization within living cells.
最近对转录组的分析表明，RNA执行着编码蛋白质之外的众多功能。RNA功能的关键在于其运输到独特的亚细胞位置。尽管RNA定位很重要，但以无偏差的方式进行研究仍然极具挑战性。我们最近描述了通过空间限制的核碱基氧化在亚细胞位置标记RNA分子的能力。在此，我们通过蛋白质的局部氧化和标记描述了该方案的显著改进。RNA-蛋白质复合物的分离使得在染色质上富集具有挑战性的RNA靶标成为可能，并为分析活细胞内RNA亚细胞定位提供了一个经过大幅优化的方案。
