Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets. Author Yongdae Shin, Joel Berry, Nicole Pannucci, Mikko Haataja, Jared Toettcher, Clifford Brangwynne Publication Year 2017 Type Journal Article 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 membrane-less 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 that 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 that undergo aging into irreversible aggregates. This system can thus elucidate not only physiological phase transitions but also their link to pathological aggregates. Keywords Animals, Mice, Kinetics, Molecular Imaging, Light, Proteins, NIH 3T3 Cells, Optogenetics, Arabidopsis Proteins, Protein Interaction Maps, Phase Transition, Models, Chemical, Cryptochromes, Intrinsically Disordered Proteins Journal Cell Volume 168 Issue 1-2 Pages 159-171.e14 Date Published 2017 Jan 12 ISSN Number 1097-4172 DOI 10.1016/j.cell.2016.11.054 Alternate Journal Cell PMCID PMC5562165 PMID 28041848 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML