验证应激反应长链非编码 RNA 与 RNA 结合蛋白在体外的相互作用。

Validation of the Interaction Between Stress-Responsive LncRNAs and RNA-Binding Proteins In Vitro.

机构信息

College of Grassland Agriculture, Northwest A&F University, Yangling, China.

Department of Biology, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China.

出版信息

Methods Mol Biol. 2024;2832:133-144. doi: 10.1007/978-1-0716-3973-3_9.

DOI:10.1007/978-1-0716-3973-3_9

PMID:38869792

Abstract

Plant long non-coding RNAs (lncRNAs) have been implicated in many biological processes, including responses to abiotic stresses, yet their detailed functions and especially their modes of action are still underexplored. lncRNAs often interact with proteins to participate in multiple levels of gene regulation. Therefore, identifying the RNA-binding proteins and validating their interaction with lncRNAs will be instrumental in revealing the functions of lncRNAs. Here, we describe two major methods to determine the interaction between lncRNA and proteins in vitro, RNA pull-down, and RNA EMSA.

摘要

植物长非编码 RNA（lncRNAs）被认为参与了许多生物过程，包括对非生物胁迫的反应，但它们的详细功能，特别是它们的作用模式，仍未得到充分探索。lncRNAs 通常与蛋白质相互作用，参与多个层次的基因调控。因此，鉴定 RNA 结合蛋白并验证其与 lncRNA 的相互作用，对于揭示 lncRNA 的功能至关重要。在这里，我们描述了两种主要的方法来确定 lncRNA 与蛋白质在体外的相互作用，RNA 下拉和 RNA EMSA。