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Identification of pre-miRNA156a binding proteins by MCP-MS2/TurboID

ABSTRACT: This study developed an integrated MCP-MS2/TurboID proximity labeling system to identify pre-miRNA156a-interacting proteins in vitro. By co-expressing MS2-tagged pre-miRNA156a with MCP-TurboID fusion protein, we achieved specific recruitment of TurboID to the target RNA. Following a 10-minute biotin pulse, proximal proteins were biotinylated, enriched using streptavidin magnetic beads, and separated by SDS-PAGE. Comparative analysis of differential protein bands between experimental and control groups, followed by LC-MS/MS analysis of excised gel bands (35-55 kDa), enabled systematic identification of pre-miRNA156a-binding proteins. This approach combines the specificity of MCP-MS2 interaction with the sensitivity of TurboID-mediated proximity labeling for comprehensive RNA-protein interactome profiling.

ORGANISM(S): Arabidopsis Thaliana

SUBMITTER: Yunfeng Liu

PROVIDER: PXD066141 | iProX | Tue Jul 15 00:00:00 BST 2025

REPOSITORIES: iProX

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