Designer membraneless organelles sequester native factors for control of cell behavior

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. 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 90% of 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.

Automated summary

The subcellular compartmentalization of macromolecules is essential for controlling biochemical reactions by increasing flux and preventing inhibitory interactions. We have developed a synthetic membraneless organelle platform that can control endogenous cellular activities by sequestering and insulating native proteins. By relocalizing targeted enzymes to synthetic condensates, we are able to efficiently regulate cellular behaviors such as proliferation, division, and cytoskeletal organization, and demonstrate strategies for releasing cargo from condensates to switch cells between functional states. These synthetic organelles offer a powerful and versatile approach to controlling cell decision-making in various model systems, with broad applications in cellular engineering.