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Probing and engineering liquid-phase organelles

Nature Biotechnology volume 37, pages 1435–1445 (2019)Cite this article

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Abstract

Cells compartmentalize their intracellular environment to orchestrate countless simultaneous biochemical processes. Many intracellular tasks rely on membrane-less organelles, multicomponent condensates that assemble by liquid–liquid phase separation. A decade of intensive research has provided a basic understanding of the biomolecular driving forces underlying the form and function of such organelles. Here we review the technologies enabling these developments, along with approaches to designing spatiotemporally actuated organelles based on multivalent low-affinity interactions. With these recent advances, it is now becoming possible both to modulate the properties of native condensates and to engineer entirely new structures, with the potential for widespread biomedical and biotechnological applications.

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Fig. 1: Diversity of condensate forms and functions in living cells.
Fig. 2: Techniques and technologies for interrogating membrane-less organelles.
Fig. 3: Multivalency in native and engineered condensates.
Fig. 4: Applications of engineered condensates.

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References

  1. Brangwynne, C. P. et al. Germline P granules are liquid droplets that localize by controlled dissolution/condensation. Science 324, 1729–1732 (2009).

    Article  CAS  PubMed  Google Scholar 

  2. Shin, Y. & Brangwynne, C. P. Liquid phase condensation in cell physiology and disease. Science 357, eaaf4382 (2017).

    Article  PubMed  CAS  Google Scholar 

  3. Banani, S. F., Lee, H. O., Hyman, A. A. & Rosen, M. K. Biomolecular condensates: organizers of cellular biochemistry. Nat. Rev. Mol. Cell Biol. 18, 285–298 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Berry, J., Weber, S. C., Vaidya, N., Haataja, M. & Brangwynne, C. P. RNA transcription modulates phase transition-driven nuclear body assembly. Proc. Natl Acad. Sci. USA 112, E5237–E5245 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Molliex, A. et al. Phase separation by low complexity domains promotes stress granule assembly and drives pathological fibrillization. Cell 163, 123–133 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Sabari, B. R. et al. Coactivator condensation at super-enhancers links phase separation and gene control. Science 361, eaar3958 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  7. Strom, A. R. et al. Phase separation drives heterochromatin domain formation. Nature 547, 241–245 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Larson, A. G. et al. Liquid droplet formation by HP1α suggests a role for phase separation in heterochromatin. Nature 547, 236–240 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Shin, Y. et al. Liquid nuclear condensates mechanically sense and restructure the genome. Cell 175, 1481–1491.e13 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Cho, W.-K. et al. Mediator and RNA polymerase II clusters associate in transcription-dependent condensates. Science 361, 412–415 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Flory, P. J. Principles of Polymer Chemistry (Cornell University Press, 1953).

  12. Rubinstein, M. & Colby, R. H. Polymer Physics (Oxford University Press, 2003).

  13. Li, P. et al. Phase transitions in the assembly of multivalent signalling proteins. Nature 483, 336–340 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Brangwynne, C. P., Tompa, P. & Pappu, R. V. Polymer physics of intracellular phase transitions. Nat. Phys. 11, 899–904 (2015).

    Article  CAS  Google Scholar 

  15. Putnam, A., Cassani, M., Smith, J. & Seydoux, G. A gel phase promotes condensation of liquid P granules in Caenorhabditis elegans embryos. Nat. Struct. Mol. Biol. 26, 220–226 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Roux, K. J., Kim, D. I., Raida, M. & Burke, B. A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells. J. Cell Biol. 196, 801–810 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Rhee, H. W. et al. Proteomic mapping of mitochondria in living cells via spatially restricted enzymatic tagging. Science 339, 1328–1331 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Markmiller, S. et al. Context-dependent and disease-specific diversity in protein interactions within stress granules. Cell 172, 590–604.e13 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Ramanathan, M. et al. RNA-protein interaction detection in living cells. Nat. Methods 15, 207–212 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Huang, R., Han, M., Meng, L. & Chen, X. Transcriptome-wide discovery of coding and noncoding RNA-binding proteins. Proc. Natl Acad. Sci. USA 115, E3879–E3887 (2018).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Schmidtmann, E., Anton, T., Rombaut, P., Herzog, F. & Leonhardt, H. Determination of local chromatin composition by CasID. Nucleus 7, 476–484 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Myers, S. A. et al. Discovery of proteins associated with a predefined genomic locus via dCas9-APEX-mediated proximity labeling. Nat. Methods 15, 437–439 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Gao, X. D. et al. C-BERST: defining subnuclear proteomic landscapes at genomic elements with dCas9-APEX2. Nat. Methods 15, 433–436 (2018).

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Quinodoz, S. A. et al. Higher-order inter-chromosomal hubs shape 3D genome organization in the nucleus. Cell 174, 744–757.e24 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Klar, T. A., Jakobs, S., Dyba, M., Egner, A. & Hell, S. W. Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission. Proc. Natl Acad. Sci. USA 97, 8206–8210 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Betzig, E. et al. Imaging intracellular fluorescent proteins at nanometer resolution. Science 313, 1642–1645 (2006).

    Article  CAS  PubMed  Google Scholar 

  27. Rust, M. J., Bates, M. & Zhuang, X. Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat. Methods 3, 793–795 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Wang, J. T. et al. Regulation of RNA granule dynamics by phosphorylation of serine-rich, intrinsically disordered proteins in C. elegans. eLife 3, e04591 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

  29. Jain, S. et al. ATPase-modulated stress granules contain a diverse proteome and substructure. Cell 164, 487–498 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. West, J. A. et al. Structural, super-resolution microscopy analysis of paraspeckle nuclear body organization. J. Cell Biol. 214, 817–830 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Fei, J. et al. Quantitative analysis of multilayer organization of proteins and RNA in nuclear speckles at super resolution. J. Cell. Sci. 130, 4180–4192 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Asano, S., Engel, B. D. & Baumeister, W. In situ cryo-electron tomography: a post-reductionist approach to structural biology. J. Mol. Biol. 428, 332–343 (2016). 2 Pt A.

    Article  CAS  PubMed  Google Scholar 

  33. Freeman Rosenzweig, E. S. et al. The eukaryotic CO2-concentrating organelle is liquid-like and exhibits dynamic reorganization. Cell 171, 148–162.e19 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Delarue, M. et al. mTORC1 controls phase separation and the biophysical properties of the cytoplasm by tuning crowding. Cell 174, 338–349.e20 (2018).

    Article  CAS  PubMed  Google Scholar 

  35. Guo, Q. et al. In Situ Structure of Neuronal C9orf72 Poly-GA Aggregates Reveals Proteasome Recruitment. Cell 172, 696–705.e12 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Ishikawa-Ankerhold, H. C., Ankerhold, R. & Drummen, G. P. C. Advanced fluorescence microscopy techniques—FRAP, FLIP, FLAP, FRET and FLIM. Molecules 17, 4047–4132 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Mueller, F., Mazza, D., Stasevich, T. J. & McNally, J. G. FRAP and kinetic modeling in the analysis of nuclear protein dynamics: what do we really know? Curr. Opin. Cell Biol. 22, 403–411 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Taylor, N. O., Wei, M., Stone, H. A. & Brangwynne, C. P. Quantifying phase-separated protein condensates using fluorescence recovery after photobleaching. Biophys. J. 117, 1285–1300 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Weidtkamp-Peters, S. et al. Dynamics of component exchange at PML nuclear bodies. J. Cell Sci. 121, 2731–2743 (2008).

    Article  CAS  PubMed  Google Scholar 

  40. Magde, D., Elson, E. & Webb, W. W. Thermodynamic fluctuations in a reacting system—measurement by fluorescence correlation spectroscopy. Phys. Rev. Lett. 29, 705–708 (1972).

    Article  CAS  Google Scholar 

  41. Wei, M.-T. et al. Phase behaviour of disordered proteins underlying low density and high permeability of liquid organelles. Nat. Chem. 9, 1118–1125 (2017).

    Article  CAS  PubMed  Google Scholar 

  42. Bracha, D. et al. Mapping local and global liquid phase behavior in living cells using photo-oligomerizable seeds. Cell 175, 1467–1480.e13 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Pliss, A. et al. Cycles of protein condensation and discharge in nuclear organelles studied by fluorescence lifetime imaging. Nat. Commun. 10, 455 (2019).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  44. Kaminski Schierle, G. S. et al. A FRET sensor for non-invasive imaging of amyloid formation in vivo. ChemPhysChem 12, 673–680 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Laine, R. F. et al. Fast fluorescence lifetime imaging reveals the aggregation processes of α-synuclein and polyglutamine in aging Caenorhabditis elegans. ACS Chem. Biol. 14, 1628–1636 (2019).

    Article  CAS  PubMed  Google Scholar 

  46. Gall, J. G., Bellini, M., Wu, Z. & Murphy, C. Assembly of the nuclear transcription and processing machinery: Cajal bodies (coiled bodies) and transcriptosomes. Mol. Biol. Cell 10, 4385–4402 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Feric, M. et al. Coexisting liquid phases underlie nucleolar subcompartments. Cell 165, 1686–1697 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Brangwynne, C. P., Mitchison, T. J. & Hyman, A. A. Active liquid-like behavior of nucleoli determines their size and shape in Xenopus laevis oocytes. Proc. Natl Acad. Sci. USA 108, 4334–4339 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Feric, M. & Brangwynne, C. P. A nuclear F-actin scaffold stabilizes ribonucleoprotein droplets against gravity in large cells. Nat. Cell Biol. 15, 1253–1259 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Eggers, J., Lister, J. R. & Stone, H. A. Coalescence of liquid drops. J. Fluid Mech. 401, 293–310 (1999).

    Article  CAS  Google Scholar 

  51. Elbaum-Garfinkle, S. et al. The disordered P granule protein LAF-1 drives phase separation into droplets with tunable viscosity and dynamics. Proc. Natl Acad. Sci. USA 112, 7189–7194 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Caragine, C. M., Haley, S. C. & Zidovska, A. Surface fluctuations and coalescence of nucleolar droplets in the human cell nucleus. Phys. Rev. Lett. 121, 148101 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Hubstenberger, A., Noble, S. L., Cameron, C. & Evans, T. C. Translation repressors, an RNA helicase, and developmental cues control RNP phase transitions during early development. Dev. Cell 27, 161–173 (2013).

    Article  CAS  PubMed  Google Scholar 

  54. Patel, A. et al. ATP as a biological hydrotrope. Science 356, 753–756 (2017).

    Article  CAS  PubMed  Google Scholar 

  55. Wirtz, D. Particle-tracking microrheology of living cells: principles and applications. Annu. Rev. Biophys. 38, 301–326 (2009).

    Article  CAS  PubMed  Google Scholar 

  56. Mason, T. G. & Weitz, D. A. Optical measurements of frequency-dependent linear viscoelastic moduli of complex fluids. Phys. Rev. Lett. 74, 1250–1253 (1995).

    Article  CAS  PubMed  Google Scholar 

  57. Taylor, N. et al. Biophysical characterization of organelle-based RNA/protein liquid phases using microfluidics. Soft Matter 12, 9142–9150 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Jawerth, L. M. et al. Salt-dependent rheology and surface tension of protein condensates using optical traps. Phys. Rev. Lett. 121, 258101 (2018).

    Article  CAS  PubMed  Google Scholar 

  59. Alberti, S., Gladfelter, A. & Mittag, T. Considerations and challenges in studying liquid-liquid phase separation and biomolecular condensates. Cell 176, 419–434 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Castello, A. et al. Insights into RNA biology from an atlas of mammalian mRNA-binding proteins. Cell 149, 1393–1406 (2012).

    Article  CAS  PubMed  Google Scholar 

  61. Pawson, T. & Nash, P. Assembly of cell regulatory systems through protein interaction domains. Science 300, 445–452 (2003).

    Article  CAS  PubMed  Google Scholar 

  62. Cohen, G. B., Ren, R. & Baltimore, D. Modular binding domains in signal transduction proteins. Cell 80, 237–248 (1995).

    Article  CAS  PubMed  Google Scholar 

  63. Schwarz-Romond, T., Merrifield, C., Nichols, B. J. & Bienz, M. The Wnt signalling effector Dishevelled forms dynamic protein assemblies rather than stable associations with cytoplasmic vesicles. J. Cell Sci. 118, 5269–5277 (2005).

    Article  CAS  PubMed  Google Scholar 

  64. Burke, K. A., Janke, A. M., Rhine, C. L. & Fawzi, N. L. Residue-by-residue view of in vitro FUS granules that bind the C-terminal domain of RNA polymerase II.

  65. Monahan, Z. et al. Phosphorylation of the FUS low-complexity domain disrupts phase separation, aggregation, and toxicity. EMBO J. 36, 2951–2967 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. Brady, J. P. et al. Structural and hydrodynamic properties of an intrinsically disordered region of a germ cell-specific protein on phase separation. Proc. Natl Acad. Sci. USA 114, E8194–E8203 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  67. Nott, T. J. et al. Phase transition of a disordered nuage protein generates environmentally responsive membraneless organelles. Mol. Cell 57, 936–947 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Wang, J. et al. A Molecular grammar governing the driving forces for phase separation of prion-like RNA binding proteins. Cell 174, 688–699.e16 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  69. Riback, J. A. et al. Stress-triggered phase separation is an adaptive, evolutionarily tuned response. Cell 168, 1028–1040.e19 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Das, R. K. & Pappu, R. V. Conformations of intrinsically disordered proteins are influenced by linear sequence distributions of oppositely charged residues. Proc. Natl Acad. Sci. USA 110, 13392–13397 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  71. Martin, E. W. & Mittag, T. Relationship of sequence and phase separation in protein low-complexity regions. Biochemistry 57, 2478–2487 (2018).

    Article  CAS  PubMed  Google Scholar 

  72. Simon, J. R., Carroll, N. J., Rubinstein, M., Chilkoti, A. & López, G. P. Programming molecular self-assembly of intrinsically disordered proteins containing sequences of low complexity. Nat. Chem. 9, 509–515 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Roberts, S., Dzuricky, M. & Chilkoti, A. Elastin-like polypeptides as models of intrinsically disordered proteins. FEBS Lett. 589, 2477–2486 (2015). 19 Pt A.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  74. Urry, D. W. Physical chemistry of biological free energy transduction as demonstrated by elastic protein-based polymers †. J. Phys. Chem. B 101, 11007–11028 (1997).

    Article  CAS  Google Scholar 

  75. Urry, D. W. The change in Gibbs free energy for hydrophobic association: Derivation and evaluation by means of inverse temperature transitions. Chem. Phys. Lett. 399, 177–183 (2004).

    CAS  Google Scholar 

  76. Quiroz, F. G. & Chilkoti, A. Sequence heuristics to encode phase behaviour in intrinsically disordered protein polymers. Nat. Mater. 14, 1164–1171 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Banani, S. F. et al. Compositional control of phase-separated cellular bodies. Cell 166, 651–663 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  78. Schuster, B. S. et al. Controllable protein phase separation and modular recruitment to form responsive membraneless organelles. Nat. Commun. 9, 2985 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  79. Nott, T. J., Craggs, T. D. & Baldwin, A. J. Membraneless organelles can melt nucleic acid duplexes and act as biomolecular filters. Nat. Chem. 8, 569–575 (2016).

    Article  CAS  PubMed  Google Scholar 

  80. Shin, Y. et al. Spatiotemporal control of intracellular phase transitions using light-activated optoDroplets. Cell 168, 159–171.e14 (2017).

    Article  CAS  PubMed  Google Scholar 

  81. Más, P., Devlin, P. F., Panda, S. & Kay, S. A. Functional interaction of phytochrome B and cryptochrome 2. Nature 408, 207–211 (2000).

    Article  PubMed  Google Scholar 

  82. Zhang, P. et al. Chronic optogenetic induction of stress granules is cytotoxic and reveals the evolution of ALS-FTD pathology. eLife 8, e39578 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  83. Wei, M.-T., Chang, Y.-C., Shimobayashi, S. F., Shin, Y. & Brangwynne, C. P. Nucleated transcriptional condensates amplify gene expression. Preprint at https://www.biorxiv.org/content/10.1101/737387 (2019).

  84. Liao, Y.-C. et al. RNA granules hitchhike on lysosomes for long-distance transport, using annexin A11 as a molecular tether. SSRN Electron. J. https://doi.org/10.2139/ssrn.3312723 (2019).

  85. Mann, J. R. et al. RNA binding antagonizes neurotoxic phase transitions of TDP-43. Neuron 102, 321–338.e8 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  86. Taslimi, A. et al. An optimized optogenetic clustering tool for probing protein interaction and function. Nat. Commun. 5, 4925 (2014).

    Article  CAS  PubMed  Google Scholar 

  87. Zhu, L. et al. Controlling the material properties and rRNA processing function of the nucleolus using light. Proc. Natl Acad. Sci. USA 116, 17330–17335 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Dine, E., Gil, A. A., Uribe, G., Brangwynne, C. P. & Toettcher, J. E. Protein phase separation provides long-term memory of transient spatial stimuli. Cell Syst. 6, 655–663.e5 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  89. Nakamura, H. et al. Intracellular production of hydrogels and synthetic RNA granules by multivalent molecular interactions. Nat. Mater. 17, 79–89 (2018).

    Article  CAS  PubMed  Google Scholar 

  90. Chong, S. et al. Imaging dynamic and selective low-complexity domain interactions that control gene transcription. Science 361, eaar2555 (2018).

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  91. Zwicker, D., Seyboldt, R., Weber, C. A., Hyman, A. A. & Jülicher, F. Growth and division of active droplets provides a model for protocells. Nat. Phys. 13, 408–413 (2017).

    Article  CAS  Google Scholar 

  92. Strulson, C. A., Molden, R. C., Keating, C. D. & Bevilacqua, P. C. RNA catalysis through compartmentalization. Nat. Chem. 4, 941–946 (2012).

    Article  CAS  PubMed  Google Scholar 

  93. Drobot, B. et al. Compartmentalised RNA catalysis in membrane-free coacervate protocells. Nat. Commun. 9, 3643 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  94. Wang, H. et al. CRISPR-mediated programmable 3D genome positioning and nuclear organization. Cell 175, 1405–1417.e14 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  95. Rege, M. et al. LADL: light-activated dynamic looping for endogenous gene expression control. Preprint at https://www.biorxiv.org/content/10.1101/349340 (2018).

  96. Hammer, S. K. & Avalos, J. L. Harnessing yeast organelles for metabolic engineering. Nat. Chem. Biol. 13, 823–832 (2017).

    Article  CAS  PubMed  Google Scholar 

  97. Castellana, M. et al. Enzyme clustering accelerates processing of intermediates through metabolic channeling. Nat. Biotechnol. 32, 1011–1018 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  98. An, S., Kumar, R., Sheets, E. D. & Benkovic, S. J. Reversible compartmentalization of de novo purine biosynthetic complexes in living cells. Science 320, 103–106 (2008).

    Article  CAS  PubMed  Google Scholar 

  99. Zhao, E. M. et al. Light-based control of metabolic flux through assembly of synthetic organelles. Nat. Chem. Biol. 15, 589–597 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  100. Woodruff, J. B. et al. The centrosome is a selective condensate that nucleates microtubules by concentrating tubulin. Cell 169, 1066–1077.e10 (2017).

    Article  CAS  PubMed  Google Scholar 

  101. Su, X. et al. Phase separation of signaling molecules promotes T cell receptor signal transduction. Science 352, 595–599 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  102. Reinkemeier, C. D., Girona, G. E. & Lemke, E. A. Designer membraneless organelles enable codon reassignment of selected mRNAs in eukaryotes. Science 363, eaaw2644 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  103. McDaniel, J. R., Callahan, D. J. & Chilkoti, A. Drug delivery to solid tumors by elastin-like polypeptides. Adv. Drug Deliv. Rev. 62, 1456–1467 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  104. Mullard, A. Biomolecular condensates pique drug discovery curiosity. Nat. Rev. Drug Discov. https://doi.org/10.1038/d41573-019-00069-w (2019).

  105. Case, L. B., Zhang, X., Ditlev, J. A. & Rosen, M. K. Stoichiometry controls activity of phase-separated clusters of actin signaling proteins. Science 363, 1093–1097 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  106. Boke, E. et al. Amyloid-like self-assembly of a cellular compartment. Cell 166, 637–650 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  107. Mateju, D. et al. An aberrant phase transition of stress granules triggered by misfolded protein and prevented by chaperone function. EMBO J. 36, 1669–1687 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  108. Jastrow, H. Dr. Jastrow’s Electron Microscopic Atlas http://www.drjastrow.de/WAI/EM/EMAtlas.html (accessed 23 October 2019).

  109. Rubinstein, M. & Colby, R. H. Polymer Physics (Oxford University Press, 2003).

  110. Berry, J., Brangwynne, C. P. & Haataja, M. Physical principles of intracellular organization via active and passive phase transitions. Rep. Prog. Phys. 81, 046601 (2018).

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

We thank members of the Brangwynne laboratory for comments on the manuscript. This work was supported by the Howard Hughes Medical Institute and by grants from the NIH 4D Nucleome Program (U01 DA040601), the Princeton Center for Complex Materials, and an NSF-supported MRSEC (DMR 1420541). D.B. acknowledges support through a Cross-Disciplinary Postdoctoral Fellowship from the Human Frontiers Science Program. M.T.W. is supported by the National Science Foundation Graduate Research Fellowship Program under grant no. DGE-1656466.

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  1. These authors contributed equally: Dan Bracha, Mackenzie T. Walls.

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  1. Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ, USA

    Dan Bracha, Mackenzie T. Walls & Clifford P. Brangwynne

  2. Howard Hughes Medical Institute, Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ, USA

    Clifford P. Brangwynne

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Bracha, D., Walls, M.T. & Brangwynne, C.P. Probing and engineering liquid-phase organelles. Nat Biotechnol 37, 1435–1445 (2019). https://doi.org/10.1038/s41587-019-0341-6

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