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Protocol for detecting lncRNA-protein interactions in vitro by tRSA RNA pull-down assay.


ABSTRACT: Long non-coding RNAs (lncRNAs) work together with diverse RNA-binding proteins (RBPs) to fulfill key regulations in important cellular functions. Here, we present a protocol to detect lncRNA-RBP interactions in vitro using a tRNA scaffold containing a streptavidin aptamer pull-down assay. We describe steps for preparing both protein and lncRNA transcripts, lncRNA-protein interaction detection with an in vitro binding assay, and western blot analysis. This protocol is applicable to screen for RNA-interacting proteins using cell lysates followed by mass spectrometry analysis. For complete details on the use and execution of this protocol, please refer to Yang et al. (2023).1.

SUBMITTER: Jiang L 

PROVIDER: S-EPMC10789616 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature

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Protocol for detecting lncRNA-protein interactions in vitro by tRSA RNA pull-down assay.

Jiang Liyun L   Yang Jun J   He Reqing R   Zhu Youlin Y   Wang Dong D  

STAR protocols 20240105 1


Long non-coding RNAs (lncRNAs) work together with diverse RNA-binding proteins (RBPs) to fulfill key regulations in important cellular functions. Here, we present a protocol to detect lncRNA-RBP interactions in vitro using a tRNA scaffold containing a streptavidin aptamer pull-down assay. We describe steps for preparing both protein and lncRNA transcripts, lncRNA-protein interaction detection with an in vitro binding assay, and western blot analysis. This protocol is applicable to screen for RNA  ...[more]

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