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使用小 RNA 定量测定 mRNA 的合成和降解速率。

Quantifying mRNA synthesis and decay rates using small RNAs.

机构信息

Department of Physics, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA.

出版信息

Biophys J. 2010 Jun 16;98(12):2780-4. doi: 10.1016/j.bpj.2010.03.022.

Abstract

Regulation of mRNA decay is a critical component of global cellular adaptation to changing environments. The corresponding changes in mRNA lifetimes can be coordinated with changes in mRNA transcription rates to fine-tune gene expression. Current approaches for measuring mRNA lifetimes can give rise to secondary effects due to transcription inhibition and require separate experiments to estimate changes in mRNA transcription rates. Here, we propose an approach for simultaneous determination of changes in mRNA transcription rate and lifetime using regulatory small RNAs (sRNAs) to control mRNA decay. We analyze a stochastic model for coupled degradation of mRNAs and sRNAs and derive exact results connecting RNA lifetimes and transcription rates to mean abundances. The results obtained are then generalized to include nonstoichiometric coupled degradation of sRNAs. Our analysis suggests experimental protocols for determining parameters controlling the efficiency of stoichiometric regulation by small RNAs and for analyzing factors and processes regulating changes in mRNA transcription and decay.

摘要

mRNA 衰变的调控是细胞适应环境变化的整体过程中的一个关键组成部分。mRNA 寿命的相应变化可以与 mRNA 转录率的变化相协调,以精细调节基因表达。目前测量 mRNA 寿命的方法可能会由于转录抑制而产生次生效应,并且需要单独的实验来估计 mRNA 转录率的变化。在这里,我们提出了一种使用调控小 RNA(sRNA)来控制 mRNA 衰变以同时确定 mRNA 转录率和寿命变化的方法。我们分析了一个耦合的 mRNA 和 sRNA 降解的随机模型,并推导出将 RNA 寿命和转录率与平均丰度联系起来的精确结果。然后,将得到的结果推广到包括非化学计量耦合的 sRNA 降解。我们的分析提出了实验方案,用于确定控制小 RNA 调控效率的参数,并用于分析调节 mRNA 转录和衰变变化的因素和过程。

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