Protocol for detecting lncRNA-protein interaction in vivo using the yeast three-hybrid assay

Jingjing Cai; Reqing He; Yongdi Zhang; Pengxiang Zhang; Youlin Zhu; Dong Wang

doi:10.1016/j.xpro.2024.102856

Protocol for detecting lncRNA-protein interaction in vivo using the yeast three-hybrid assay

STAR Protoc. 2024 Mar 15;5(1):102856. doi: 10.1016/j.xpro.2024.102856. Epub 2024 Jan 28.

Authors

Jingjing Cai¹, Reqing He¹, Yongdi Zhang¹, Pengxiang Zhang¹, Youlin Zhu¹, Dong Wang²

Affiliations

¹ Key Laboratory of Molecular Biology and Gene Engineering in Jiangxi Province, College of Life Science, Nanchang University, Jiangxi 330031, China.
² Key Laboratory of Molecular Biology and Gene Engineering in Jiangxi Province, College of Life Science, Nanchang University, Jiangxi 330031, China. Electronic address: dongwang@ncu.edu.cn.

Abstract

Analyses of long non-coding RNA (lncRNA)-protein interactions provide key clues for understanding the molecular basis of lncRNA-modulated biological processes. Here, we detail a yeast three-hybrid assay to identify the lncRNA-interacting protein. We describe steps for lncRNA bait preparation, screening an RNA-binding proteins (RBPs) cDNA library, and validation of the lncRNA-RBP interaction. The assay can also be further applied to delineate the region of RBP that mediates the RNA-protein interaction. For complete details on the use and execution of this protocol, please refer to Zhang et al.¹.

Keywords: Gene Expression; Genetics; Plant sciences.

MeSH terms

Gene Library
RNA, Long Noncoding* / genetics
RNA-Binding Proteins / genetics
RNA-Binding Proteins / metabolism
Two-Hybrid System Techniques

Substances

RNA, Long Noncoding
RNA-Binding Proteins