Abstract
<h2>Summary</h2> Liquid-liquid phase separation (LLPS) is recognized as an important means of intracellular compartmentalization in living cells. However, the spontaneous formation of hierarchical biomacromolecule-condensed microcompartments <i>in vitro</i> capable of spatial organization, isolation, and modulated biological processes is still a challenge. Here, we develop a way for spatial organization of various biomacromolecules based on heterogeneous LLPS. We show that by the modulation of interfacial tension among the biomacromolecule condensates, three typical spatial configurations are successfully achieved, including nesting, partial engulfing, and petal-like multicompartmental models. Particularly, upon changes in the external environment, a reversible programmable reconstitution between the formed sub-microcompartments is realized, which then provides a technological platform for the spatial modulation of biological reactions. Overall, our results highlight opportunities for the programmable organization of multiple biomacromolecules condensates, which contributes a key step toward intramicrocompartmental spatial organization with the integration of various artificial organelles and cytoplasm-like liquid phase.
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