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bioRxiv · 10.1101/2025.07.29.667348

Protocol for capturing the RNA-binding proteome from plants using orthogonal organic phase separation

Sanchez-Camargo, V. A.·Kramer, G.·van den Burg, H. A.
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Abstract

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.

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Sanchez-Camargo, V. A., Kramer, G., van den Burg, H. A.. 2025-07-29. Protocol for capturing the RNA-binding proteome from plants using orthogonal organic phase separation. https://doi.org/10.1101/2025.07.29.667348

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