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Liquid-liquid phase separation (LLPS) of biopolymers recently emerged as a ubiquitous driving force for intracellular self-assembly. The resulting LLPS dynamics appear to orchestrate wide-ranging cellular mechanisms, from genome organization to cell division. Despite remarkable progress, studying intracellular LLPS remains challenging. In this talk, I will present our recent progress towards dissecting the LLPS of intrinsically-disordered proteins (IDPs) and its biological significance. I will highlight our two main approaches: (1) the engineering of lower and upper critical solution temperature (LCST and UCST, respectively) LLPS behavior in IDPs (Nature Materials 2015, Science Advances 2019), and (2) the probing of intracellular LLPS dynamics in mammalian skin (Science 2020). Dwelling on skin, I will explain how the biophysical lens of LLPS sheds new light into the enigmatic process of skin barrier formation. Using this system, I will also describe novel biomolecular tools to probe LLPS in tissue biology. Finally, I will discuss engineered IDPs that exhibit non-equilibrium LLPS. These behaviors differ from the established biophysical frameworks to study LLPS of IDPs, which were inspired by work on synthetic polymers that exhibit equilibrium LLPS. Overall, my goal is to inspire students and scientists at ICTP to connect with the research frontier of cellular mechanisms driven by LLPS. This nascent field offers fruitful ground for exploration of experimental and theoretical biophysical approaches.
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