Designer membraneless organelles sequester native factors for control of cell behavior

• Published in: Nature chemical biology Vol. 17; no. 9
• Main Authors: Garabedian, Mikael V., Wang, Wentao, Dabdoub, Jorge B., Tong, Michelle, Caldwell, Reese M., Benman, William, Schuster, Benjamin S., Deiters, Alexander, Good, Matthew C.
• Format: Journal Article
• Language: English
• Published: United States Nature Publishing Group 02.08.2021
• Subjects: BASIC BIOLOGICAL SCIENCES; Compartmentalization; disordered protein; INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; membraneless organelles; mesoscale protein switch; programmable behavior; RGG domain; synthetic condense
• ISSN: 1552-4450; 1552-4469

Subcellular compartmentalization of macromolecules increases flux and prevents inhibitory interactions to control biochemical reactions. Inspired by this functionality, we sought to build designer compartments that function as hubs to regulate the flow of information through cellular control systems. Here we report a synthetic membraneless organelle platform to control endogenous cellular activities through sequestration and insulation of native proteins. We engineer and express a disordered protein scaffold to assemble micron size condensates and recruit endogenous clients via genomic tagging with high-affinity dimerization motifs. By relocalizing up to ninety percent of a targeted enzymes to synthetic condensates, we efficiently control cellular behaviors, including proliferation, division, and cytoskeletal organization. Further, we demonstrate multiple strategies for controlled cargo release from condensates to switch cells between functional states. These synthetic organelles offer a powerful and generalizable approach to modularly control cell decision-making in a variety of model systems with broad applications for cellular engineering.