Quantitative Comparisons of Translation Activity by Ribosome Profiling with Internal Standards.
Quantitative Comparisons of Translation Activity by Ribosome Profiling with Internal Standards.
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DOI：
10.1007/978-1-0716-1150-0_5
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发表时间：
2021
影响因子：
--
通讯作者：
Yinuo J. Wang;Wendy V. Gilbert
中科院分区：
文献类型：
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作者：
Yinuo J. Wang;Wendy V. Gilbert
核糖体分析是一种全基因组的方法来绘制核糖体在信使rna上的位置。在一定条件下，核糖体保护片段的丰度可以用来比较不同转录本之间的相对翻译活性，以及同一转录本在不同条件下的相对翻译活性。然而，目前缺乏一种统一的、常规的方法来规范全球翻译水平差异的条件之间的关系。在这里，我们描述了实验和计算方法，以使用正交物种尖峰，或内部标准，使翻译活动的绝对比较条件之间。这种对标准核糖体分析的简单修改为准确解释翻译的各种遗传、化学和环境扰动的影响提供了一种可靠的方法。
Ribosome profiling is a genome-wide approach to map the positions of ribosomes on messenger RNAs. The abundance of ribosome-protected fragments can be used within condition to compare relative translation activities between different transcripts and between distinct conditions for the same transcript. A unified and routine method is currently lacking, however, to normalize between conditions for differences in global translation levels. Here we describe experimental and computational methods to use an orthogonal species spike-in, or internal standard, to enable absolute comparisons of translation activity between conditions. This simple modification of standard ribosome profiling provides a robust approach for accurately interpreting the effects of diverse genetic, chemical, and environmental perturbations of translation.