Thor-Ribo-Seq: ribosome profiling tailored for low input with RNA-dependent RNA amplification
Thor-Ribo-Seq: ribosome profiling tailored for low input with RNA-dependent RNA amplification
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DOI：
10.1101/2023.01.15.524129
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发表时间：
2023-01
期刊：
影响因子：
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通讯作者：
M. Mito;Yuichi Shichino;Shintaro Iwasaki
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文献类型：
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作者：
M. Mito;Yuichi Shichino;Shintaro Iwasaki
翻译调控在基因表达的多样化和对细胞内外环境提示的反应中起着关键作用。核糖体图谱(或称核糖体序列)是一种敏感的、定量的、全面的和数据丰富的技术，可以用来研究核糖体在细胞转录组中的穿越。然而，由于图书馆准备的复杂性，低投入的应用带来了分析方面的挑战。为了克服这个问题，我们在这里开发了Thor-Ribo-Seq，一种为低输入量身定做的核糖体图谱方法。Thor-Ribo-Seq利用RNA模板RNA转录线性放大核糖体足迹，在密码子分辨率下使用有限的人工制品评估核糖体遍历。这种高度敏感的核糖体分析方法为研究珍贵样本中的翻译体提供了一个通用的选择。
Translation regulation plays a pivotal role in the diversification of gene expression and the response to intra- and extracellular environmental cues. Ribosome profiling (or Ribo-Seq) serves as a sensitive, quantitative, comprehensive, and data-rich technique to survey ribosome traversal across the cellular transcriptome. However, due to the intricacy of library preparation, applications to low input have presented analytic challenges. To overcome this issue, here we developed Thor-Ribo-Seq, a ribosome profiling method tailored for low input. Thor-Ribo-Seq harnesses RNA-templated RNA transcription to linearly amplify ribosome footprints, assessing ribosome traversal at codon resolution with limited artifacts. This highly sensitized ribosome profiling approach provides a versatile option to investigate the translatome in precious samples.