Serena Sanulli1, Geeta J Narlikar2. 1. Department of Genetics, Stanford University, Palo Alto, California. 2. Department of Biochemistry and Biophysics, UCSF, San Francisco, California.
Abstract
Liquid-liquid phase separation (LLPS) has been invoked as an underlying mechanism involved in the formation and function of several cellular membrane-less compartments. Given the explosion of studies in this field in recent years, it has become essential to converge on clear guidelines and methods to rigorously investigate LLPS and advance our understanding of this phenomenon. Here, we describe basic methods to (1) visualize droplets formed by nucleic acid binding proteins and (2) characterize the liquid-like nature of these droplets under controlled in vitro experimental conditions. We discuss the rationale behind these methods, as well as caveats and limitations. Our ultimate goal is to guide scientists interested in learning how to test for LLPS, while appreciating that the field is evolving rapidly and adjusting constantly to the growing knowledge.
Liquid-liquid phase separation (LLPS) has been invoked as an underlying mechanism involved in the formation and function of several cellular membrane-less compartments. Given the explosion of studies in this field in recent years, it has become essential to converge on clear guidelines and methods to rigorously investigate LLPS and advance our understanding of this phenomenon. Here, we describe basic methods to (1) visualize droplets formed by nucleic acid binding proteins and (2) characterize the liquid-like nature of these droplets under controlled in vitro experimental conditions. We discuss the rationale behind these methods, as well as caveats and limitations. Our ultimate goal is to guide scientists interested in learning how to test for LLPS, while appreciating that the field is evolving rapidly and adjusting constantly to the growing knowledge.
Authors: Amy R Strom; Alexander V Emelyanov; Mustafa Mir; Dmitry V Fyodorov; Xavier Darzacq; Gary H Karpen Journal: Nature Date: 2017-06-21 Impact factor: 49.962
Authors: Clifford P Brangwynne; Christian R Eckmann; David S Courson; Agata Rybarska; Carsten Hoege; Jöbin Gharakhani; Frank Jülicher; Anthony A Hyman Journal: Science Date: 2009-05-21 Impact factor: 47.728
Authors: Shana Elbaum-Garfinkle; Younghoon Kim; Krzysztof Szczepaniak; Carlos Chih-Hsiung Chen; Christian R Eckmann; Sua Myong; Clifford P Brangwynne Journal: Proc Natl Acad Sci U S A Date: 2015-05-26 Impact factor: 11.205
Authors: Joshua A Riback; Lian Zhu; Mylene C Ferrolino; Michele Tolbert; Diana M Mitrea; David W Sanders; Ming-Tzo Wei; Richard W Kriwacki; Clifford P Brangwynne Journal: Nature Date: 2020-05-06 Impact factor: 49.962
Authors: Diana M Mitrea; Bappaditya Chandra; Mylene C Ferrolino; Eric B Gibbs; Michele Tolbert; Michael R White; Richard W Kriwacki Journal: J Mol Biol Date: 2018-07-24 Impact factor: 5.469
Authors: Marina Feric; Nilesh Vaidya; Tyler S Harmon; Diana M Mitrea; Lian Zhu; Tiffany M Richardson; Richard W Kriwacki; Rohit V Pappu; Clifford P Brangwynne Journal: Cell Date: 2016-05-19 Impact factor: 41.582
Authors: Masato Kato; Tina W Han; Shanhai Xie; Kevin Shi; Xinlin Du; Leeju C Wu; Hamid Mirzaei; Elizabeth J Goldsmith; Jamie Longgood; Jimin Pei; Nick V Grishin; Douglas E Frantz; Jay W Schneider; She Chen; Lin Li; Michael R Sawaya; David Eisenberg; Robert Tycko; Steven L McKnight Journal: Cell Date: 2012-05-11 Impact factor: 41.582