ATP-fueled autonomous pathway-selective signal transduction on DNA-nanostructure tracks.

Chemically fueled non-equilibrium self-assemblies offer a promising route toward constructing artificial living systems, yet current strategies lack molecular-level control over where such artificial non-equilibrium assemblies occur. Here, we present an ATP-fueled enzymatic reaction network comprising DNA ligase and nickase, concatenated with a hairpin chain reaction (HCR), enabling site-confined transient covalent HCR (cHCR). Anchoring the initiator strands on paramagnetic colloids enables fuel-driven cHCRs on the colloidal surfaces, while the resulting DNA polymers gradually degrade by terminal monomer shedding upon ATP consumption. Furthermore, tethering DNA hairpins onto a DNA-nanostructure track allows for ATP-fueled transient signal transduction along predefined pathways, with multiple pathways selectively activated by their corresponding inputs. Critically, the integration of an RNA input with an RNase H reaction network establishes an autonomous input clearance mechanism, enabling repeated recognition and directional transduction of multiple inputs. This strategy for site-confined spatiotemporal regulation of self-assembly paves the way toward non-equilibrium nanodevices capable of programmable information processing and computation.