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Characterization of RNA content in individual phase-separated coacervate microdroplets

View ORCID ProfileDamian Wollny, Benjamin Vernot, Jie Wang, Maria Hondele, Anthony Hyman, Karsten Weis, J. Gray Camp, T.-Y. Dora Tang, Barbara Treutlein
doi: https://doi.org/10.1101/2021.03.08.434405
Damian Wollny
1Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany
7Present address: RNA Bioinformatics and High Throughput Analysis, Friedrich Schiller University Jena, Germany
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  • ORCID record for Damian Wollny
  • For correspondence: damian.wollny@uni-jena.de tang@mpi-cbg.de barbara.treutlein@bsse.ethz.ch
Benjamin Vernot
1Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany
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Jie Wang
2Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
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Maria Hondele
4Institute of Biochemistry, ETH Zurich, Zurich, Switzerland
8Present address: Biozentrum, University of Basel, Basel, Switzerland
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Anthony Hyman
2Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
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Karsten Weis
4Institute of Biochemistry, ETH Zurich, Zurich, Switzerland
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J. Gray Camp
1Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany
5Institute of Molecular and Clinical Ophthalmology, Basel, Switzerland
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T.-Y. Dora Tang
2Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
3Cluster of Excellence Physics of Life, TU Dresden, Dresden, Germany
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  • For correspondence: damian.wollny@uni-jena.de tang@mpi-cbg.de barbara.treutlein@bsse.ethz.ch
Barbara Treutlein
1Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany
6Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland
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  • For correspondence: damian.wollny@uni-jena.de tang@mpi-cbg.de barbara.treutlein@bsse.ethz.ch
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Abstract

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.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted March 08, 2021.
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Characterization of RNA content in individual phase-separated coacervate microdroplets
Damian Wollny, Benjamin Vernot, Jie Wang, Maria Hondele, Anthony Hyman, Karsten Weis, J. Gray Camp, T.-Y. Dora Tang, Barbara Treutlein
bioRxiv 2021.03.08.434405; doi: https://doi.org/10.1101/2021.03.08.434405
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Characterization of RNA content in individual phase-separated coacervate microdroplets
Damian Wollny, Benjamin Vernot, Jie Wang, Maria Hondele, Anthony Hyman, Karsten Weis, J. Gray Camp, T.-Y. Dora Tang, Barbara Treutlein
bioRxiv 2021.03.08.434405; doi: https://doi.org/10.1101/2021.03.08.434405

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