Sustained, Reversible, and Adaptive Non‐Equilibrium Steady States of a Dissipative DNA‐Based System

Abstract Inspired by nature, researchers have developed several chemical fuel‐driven supramolecular systems aimed at achieving improved kinetic control over their formation and functions. Alongside, DNA‐based systems regulated by energy‐dissipating mechanisms have been reported. However, the majority of these systems rely on batchwise additions of chemical fuels to closed reactors, resulting in transient non‐equilibrium states that differ fundamentally from the sustained and highly adaptable non‐equilibrium steady states (NESS) maintained by living systems through continuous energy dissipation. Here, we demonstrate sustained NESS of a dissipative DNA strand‐displacement reaction achieved through the continuous supply of an RNA fuel to an open semi‐batch reactor, using a custom automated setup that enables tunable fuel infusion rates and in situ analysis. Similar to biological NESS, our system dynamically adapts in real‐time to subtle variations in fuel supply, achieving different steady‐state levels of the strand‐displacement reaction. Our approach demonstrates remarkable on‐the‐fly control over a dissipative DNA nanosystem, unachievable when working under batch conditions. Importantly, by fitting the experimental data to a kinetic model of the reaction network, we were able to confirm that the observed steady states correspond to true non‐equilibrium compositions of the system.