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Capturing the RNA-binding proteome from plants using orthogonal organic phase separation

ABSTRACT: Summary RNA-binding proteins (RBPs) regulate many processes related to RNA biogenesis, localization, half-life and function. Here, we present a protocol for the en masse isolation of RBPs cross-linked to RNA and provide a strategy for proteomics analysis. We describe the steps for in vivo UV-crosslinking of RNA-protein complexes, tissue lysis, fractionation and purification of crosslinked RNA-protein adducts using organic solvents. Although the protocol was developed for Nicotiana benthamiana leaves, it can be optimized for use in different plants and tissues.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Nicotiana Benthamiana (ncbitaxon:4100)

SUBMITTER: Gertjan

PROVIDER: MSV000099169 | MassIVE |

SECONDARY ACCESSION(S): PXD068386

REPOSITORIES: MassIVE

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