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Characterization of RNA content in individual phase-separated coacervate microdroplets

By Damian Wollny, Benjamin Vernot, Jie Wang, Maria Hondele, Anthony A Hyman, Karsten Weis, J. Gray Camp, T-Y. Dora Tang, Barbara Treutlein

Posted 08 Mar 2021
bioRxiv DOI: 10.1101/2021.03.08.434405

Liquid-liquid phase separation or condensation is a form of macromolecular compartmentalization. Condensates formed by complex coacervation were hypothesized to have played a crucial part during the origin-of-life. In living cells, condensation organizes biomolecules into a wide range of membraneless compartments. Although RNA is a key component of condensation in cells and the central component of the RNA world hypothesis, little is known about what determines RNA accumulation in condensates and how single condensates differ in their RNA composition. Therefore, we developed an approach to read the RNA content from single condensates using high-throughput sequencing. We find that RNAs which are enriched for specific sequence motifs efficiently accumulate in condensates. These motifs show high sequence similarity to short interspersed elements (SINEs). We observed similar results for protein-derived condensates, demonstrating applicability across different in vitro reconstituted membraneless organelles. Thus, our results provide a new inroad to explore the RNA content of phase-separated droplets at single condensate resolution.

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