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Author:
Date:
2014-04-05
[Abstract] The fate of mRNA, in particular its stability, localization and rate of translation is regulated by RNA binding proteins assembling to messenger ribonucleoprotein (mRNP) complexes. To investigate the transcriptome-wide RNA binding sites of UPF1, the core factor of nonsense-mediated mRNA decay (NMD), we performed individual-nucleotide-resolution UV cross-linking and immunoprecipitation (iCLIP) (Zund et al., 2013) followed by high-throughput sequencing. The presented protocol is optimized to investigate the RNA-binding sites of UPF1 and is based on previously described studies (Konig et ...
[摘要] mRNA的命运，特别是其稳定性，定位和翻译速率由装配到信使核糖核蛋白（mRNP）复合物的RNA结合蛋白调节。 为了研究无义介导的mRNA衰变（NMD）的核心因子UPF1的转录组范围的RNA结合位点，我们进行单个核苷酸分辨率的UV交联和免疫沉淀（iCLIP）（Zund等，/em>，2013），然后进行高通量测序。 优化所提出的方案以研究UPF1的RNA结合位点并且基于先前描述的研究（Konig等人，2010; Konig等人，2011; Hafner等人，2010）。 我们要感谢Mihaela Zavolan（瑞士巴塞尔瑞士生物信息学研究所）和Jernej Ule（英国剑桥的分子生物学医学研究委员会实验室）组织这些实验的技术支持。
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