Department of Chemical Sciences and Technologies, University of Rome Tor Vergata, Via della Ricerca Scientifica, Rome 00133, Italy.
Department of Chemical Engineering and Biotechnology, University of Cambridge, Philippa Fawcett Drive, Cambridge CB3 0AS, U.K.
J Am Chem Soc. 2024 Nov 20;146(46):31529-31537. doi: 10.1021/jacs.4c08919. Epub 2024 Nov 6.
Membrane-less compartments and organelles are widely acknowledged for their role in regulating cellular processes, and there is an urgent need to harness their full potential as both structural and functional elements of synthetic cells. Despite rapid progress, synthetically recapitulating the nonequilibrium, spatially distributed responses of natural membrane-less organelles remains elusive. Here, we demonstrate that the activity of nucleic-acid cleaving enzymes can be localized within DNA-based membrane-less compartments by sequestering the respective DNA or RNA substrates. Reaction-diffusion processes lead to complex nonequilibrium patterns, dependent on enzyme concentration. By arresting similar dynamic patterns, we spatially organize different substrates in concentric subcompartments, which can be then selectively addressed by different enzymes, demonstrating spatial distribution of enzymatic activity. Besides expanding our ability to engineer advanced biomimetic functions in synthetic membrane-less organelles, our results may facilitate the deployment of DNA-based condensates as microbioreactors or platforms for the detection and quantitation of enzymes and nucleic acids.
无膜隔间和细胞器广泛用于调节细胞过程，迫切需要充分利用它们作为合成细胞的结构和功能元件的潜力。尽管取得了快速进展，但要在合成上重现天然无膜细胞器的非平衡、空间分布的响应仍然难以捉摸。在这里，我们证明可以通过隔离各自的 DNA 或 RNA 底物，将核酸切割酶的活性定位于基于 DNA 的无膜隔间内。反应-扩散过程导致复杂的非平衡模式，取决于酶浓度。通过阻止类似的动态模式，我们在同心亚隔间中空间组织不同的底物，然后可以用不同的酶选择性地处理这些底物，从而证明酶活性的空间分布。除了扩大我们在合成无膜细胞器中设计高级仿生功能的能力外，我们的结果还可能促进基于 DNA 的凝聚物作为微生物反应器或用于检测和定量酶和核酸的平台的部署。
