Programmable and Chemically Fueled DNA Coacervates by Transient Liquid-Liquid Phase Separation.

Multivalency-driven liquid-liquid phase separation (LLPS) is essential in biomolecular condensates to facilitate spatiotemporal regulation of biological functions. Providing programmable model systems would help to better understand the LLPS processes in biology, and furnish new types of compartmentalized synthetic reaction crucibles that exploit biological principles. Herein, we demonstrate a concept for programming LLPS using transient multivalency between ATP-driven sequence-defined functionalized nucleic acid polymers (SfNAPs), which serve as simple models for membrane-less organelles. The ATP-driven SfNAPs are transiently formed by an enzymatic reaction network (ERN) of concurrent ATP-powered DNA ligation and DNA restriction. The lifetimes can be programmed by the ATP concentration, which manifests on the LLPS length scale in tunable lifetimes for the all-DNA coacervates. Critically, the prominent programmability of the DNA-based building blocks allows to encode distinct molecular recognitions for multiple multivalent systems, enabling sorted LLPS and thus multicomponent DNA coacervates, reminiscent of the diverse membraneless organelles in biological systems. The ATP-driven coacervates are capable for multivalent trapping of micron-scale colloids and biomolecules to generate functions as emphasized for rate enhancements in enzymatic cascades. This work supports ATP-driven multivalent coacervation as a valuable mechanism for dynamic multicomponent and function biomolecular condensate mimics and for autonomous materials design in general.