Mechano-fluorescence actuation in single synaptic vesicles with a DNA framework nanomachine

Biomimetic machines that can convert mechanical actuation to adaptive coloration in a manner analogous to cephalopods have found widespread applications at various length scales. At the nanoscale, a transmutable nanomachine with adaptive colors that can sense and mediate cellular or intracellular interactions is highly desirable. Here, we report the design of a DNA framework nanomachine (DFN) that can autonomously change shape in response to pH variations in single synaptic vesicles, which, in turn, displays adaptive fluorescent colors with a mechano-fluorescence actuation mechanism. To construct a DFN, we used a tetrahedral DNA nanostructure as the framework to incorporate an embedded pH-responsive, i-motif sequence tagged with a Förster resonance energy transfer pair and an affinity cholesterol moiety targeting vesicular membranes. We found that endocytosed DFNs are individually trapped in single endocytic vesicles in living synaptic cells due to the size-exclusion effect. The adaptive fluorescence coloration of DFNs enabled single-vesicle quantification of resting pH values in a processive manner, allowing long-term tracking of the exocytosis and fusion dynamics in intracellular processes and cell-cell communications.