Enzyme-Responsive DNA\nCondensates

Membrane-less compartments and organelles are widely\nacknowledged\nfor their role in regulating cellular processes, and there is an urgent\nneed to harness their full potential as both structural and functional\nelements of synthetic cells. Despite rapid progress, synthetically\nrecapitulating the nonequilibrium, spatially distributed responses\nof natural membrane-less organelles remains elusive. Here, we demonstrate\nthat the activity of nucleic-acid cleaving enzymes can be localized\nwithin DNA-based membrane-less compartments by sequestering the respective\nDNA or RNA substrates. Reaction-diffusion processes lead to complex\nnonequilibrium patterns, dependent on enzyme concentration. By arresting\nsimilar dynamic patterns, we spatially organize different substrates\nin concentric subcompartments, which can be then selectively addressed\nby different enzymes, demonstrating spatial distribution of enzymatic\nactivity. Besides expanding our ability to engineer advanced biomimetic\nfunctions in synthetic membrane-less organelles, our results may facilitate\nthe deployment of DNA-based condensates as microbioreactors or platforms\nfor the detection and quantitation of enzymes and nucleic acids.