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Cell. 2019 Jan 24;176(3):419-434. doi: 10.1016/j.cell.2018.12.035.

Considerations and Challenges in Studying Liquid-Liquid Phase Separation and Biomolecular Condensates.

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Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany; Technische Universität Dresden, Center for Molecular and Cellular Bioengineering (CMCB), Biotechnology Center, 01307 Dresden, Germany. Electronic address:
University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Marine Biological Laboratory, Woods Hole, MA 02543, USA. Electronic address:
Department for Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA. Electronic address:


Evidence is now mounting that liquid-liquid phase separation (LLPS) underlies the formation of membraneless compartments in cells. This realization has motivated major efforts to delineate the function of such biomolecular condensates in normal cells and their roles in contexts ranging from development to age-related disease. There is great interest in understanding the underlying biophysical principles and the specific properties of biological condensates with the goal of bringing insights into a wide range of biological processes and systems. The explosion of physiological and pathological contexts involving LLPS requires clear standards for their study. Here, we propose guidelines for rigorous experimental characterization of LLPS processes in vitro and in cells, discuss the caveats of common experimental approaches, and point out experimental and theoretical gaps in the field.


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