Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets

Yongdae Shin, Joel Berry, Nicole Pannucci, Mikko P. Haataja, Jared E. Toettcher, Clifford P. Brangwynne

Research output: Contribution to journalArticlepeer-review

186 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)159-171.e14
JournalCell
Volume168
Issue number1-2
DOIs
StatePublished - Jan 12 2017

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Keywords

  • RNA binding protein
  • aggregation
  • fluisomes
  • gelation
  • intracellular condensates
  • intrinsically disordered protein
  • liquid-liquid phase separation
  • membraneless organelles
  • optogenetics
  • ribonucleoprotein bodies

Fingerprint Dive into the research topics of 'Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets'. Together they form a unique fingerprint.

Cite this