RNA 折叠和力下的展开：单分子实验及其分析。

RNA Folding and Unfolding Under Force: Single-Molecule Experiments and Their Analysis.

机构信息

Nanobiophysics, ESPCI Paris, Paris, France.

Department of Chemistry, University of Michigan, Ann Arbor, MI, USA.

出版信息

Methods Mol Biol. 2020;2113:89-100. doi: 10.1007/978-1-0716-0278-2_7.

DOI:10.1007/978-1-0716-0278-2_7

PMID:32006309

Abstract

We have previously described (Geffroy et al. Methods Mol Biol 1665:25-40, 2018) how to unfold (or fold) a single RNA molecule under force using a dual-beam optical trap setup. In this chapter, we complementarily describe how to analyze the corresponding data and how to interpret it in terms of RNA three-dimensional structure. As with all single-molecule methods, single RNA molecule force data often exhibit several discrete states where state-to-state transitions are blurred in a noisy signal. In order to cope with this limitation, we have implemented a novel strategy to analyze the data, which uses a hidden Markov modeling procedure. A representative example of such an analysis is presented.

摘要

我们之前曾描述过（Geffroy 等人，《方法学分子生物学》1665:25-40, 2018）如何在双光束光阱装置下使用力展开（或折叠）单个 RNA 分子。在本章中，我们补充描述了如何分析相应的数据，并如何根据 RNA 的三维结构对其进行解释。与所有单分子方法一样，单个 RNA 分子力数据通常表现出几个离散状态，其中状态到状态的转变在噪声信号中变得模糊。为了应对这一限制，我们实施了一种新的数据分析策略，该策略使用隐马尔可夫建模程序。对这种分析的一个代表性示例进行了介绍。