Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, USA.
Methods Mol Biol. 2020;2113:41-71. doi: 10.1007/978-1-0716-0278-2_5.
Data from fluorescence-based methods that measure in vivo hybridization efficacy of unique RNA regions can be used to infer regulatory activity and to identify novel RNA: RNA interactions. Here, we document the step-by-step analysis of fluorescence data collected using an in vivo regional RNA structural sensing system (iRS) for the purpose of identifying potential functional sites that are likely to be involved in regulatory interactions. We also detail a step-by-step protocol that couples this in vivo accessibility data with computational mRNA target predictions to inform the selection of potentially true targets from long lists of thermodynamic predictions.
基于荧光的方法可以测量体内独特 RNA 区域杂交效率的数据，可用于推断调控活性并鉴定新的 RNA:RNA 相互作用。在这里，我们记录了使用体内区域 RNA 结构感应系统 (iRS) 收集荧光数据的分步分析，目的是确定可能参与调控相互作用的潜在功能位点。我们还详细介绍了一种分步方案，该方案将这种体内可及性数据与计算 mRNA 靶标预测相结合，从大量热力学预测中选择潜在的真实靶标。
