EMERGE Ep 12: RNA-Protein Interactions in Space and Time скачать в хорошем качестве

EMERGE Ep 12: RNA-Protein Interactions in Space and Time

The webinar focuses on two multiomics methodologies that probe the crosstalk between the RNA and proteins, with a specific focus on characterising RNA-protein interactions and RNA and protein subcellular localisation. OOPS (Orthogonal Organic Phase Separation) repurposes standard Trizol RNA and protein extraction to unbiasedly retrieve all RNA binding proteins and protein bound RNA (Queiroz, Smith & Villanueva, Nature Biotechnology 2019; Villanueva, Smith & Queiroz, Nature Protocols 2020). The unbiased nature of OOPS allows for application across variety of human cells and the first bacterium RNA-binding proteome was also resolved. More recently, LoRNA (Localisation of RNA) was developed, a new single-step subcellular fractionation technology to quantitatively interrogate the subcellular localisation of the transcriptome, both in organelles and phase separated compartments. Simultaneous interrogation of all localisations allows the system-wide quantification of RNA proportional distribution and the comprehensive analysis of RNA subcellular dynamics. Importantly, by coupling LoRNA and LOPIT (Localisation Of Proteins by Isotope Tagging), a simultaneous system-wide map of the transcriptome and proteome could be obtained. By applying this combined framework to cells undergoing the Unfolded Protein Response the first dynamic map of how the cell coordinately reorganizes its transcriptome and proteome in stress conditions was generated with unprecedented resolution (Villanueva, Smith & Pizzinga, BioRxiv 2022).