RNA expression profiling of HEK293 cells stably expressing ARDrop-ALK

Biomolecular condensates are membraneless compartments involved in a wide range of cellular processes. Despite their fundamental role in the spatiotemporal regulation of cellular functions, tools for precisely manipulating phase-separated condensates remain limited, and effective methods for discovering and functionalizing tunable phase separation modules from natural proteins are lacking. Here we present a rational engineering approach for androgen receptor (AR) and its ligands to create a chemical genetic platform, ARDrop, enabling condensates formation and dissolution. This platform is applied to a diverse set of proteins to achieve intended cellular functions, revealing distinct signaling phases and identifying the molecular players and downstream events involved. Our work provides new insights into the dissection of complicated cell signaling landscapes and expands the toolkit for condensate manipulation, laying the foundation for engineering designer condensates for synthetic biology applications.