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Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets

Author(s): Shin, Y; Berry, J; Pannucci, N; Haataja, MP; Toettcher, JE; et al

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Abstract: Phase transitions driven by intrinsically disordered protein regions (IDRs) have emerged as a ubiquitous mechanism for assembling liquid-like RNA/protein (RNP) bodies and other membraneless organelles. However, a lack of tools to control intracellular phase transitions limits our ability to understand their role in cell physiology and disease. Here, we introduce an optogenetic platform, which uses light to activate IDR-mediated phase transitions in living cells. We use this “optoDroplet” system to study condensed phases driven by the IDRs of various RNP body proteins, including FUS, DDX4, and HNRNPA1. Above a concentration threshold, these constructs undergo light-activated phase separation, forming spatiotemporally-definable liquid optoDroplets. FUS optoDroplet assembly is fully reversible even after multiple activation cycles. However, cells driven deep within the phase boundary form solid-like gels, which undergo aging into irreversible aggregates. This system can thus elucidate not only physiological phase transitions, but also their link to pathological aggregates.
Publication Date: 12-Jan-2017
Electronic Publication Date: 12-Jan-2018
Citation: Shin, Y, Berry, J, Pannucci, N, Haataja, MP, Toettcher, JE, Brangwynne, CP. (2017). Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets. Cell, 168 (159 - 171.e14. doi:10.1016/j.cell.2016.11.054
DOI: doi:10.1016/j.cell.2016.11.054
Pages: 159 - 171
Type of Material: Journal Article
Journal/Proceeding Title: Cell
Version: Author's manuscript
Notes: Electronic publication date references PMC's acquisition and publication of the authors' manuscript.

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