Designer Dynamic DNA Nanoaggregate in Living Cell for Mitochondrial Energy Restriction

Artificial intracellular aggregations of exogenous molecules are powerful tools for modulating cellular processes and fate; however, controlling spatiotemporal assembly in a subcellular-specific manner remains challenging. In this study, a designer dynamic DNA nanoaggregate system called Tech-tetrahedron (telomerase-controllable hand-in-hand tetrahedron) based on a trinity-functionalized DNA tetrahedron with tailored vertexes was constructed to achieve spatiotemporal mitochondrial energy restriction. The navigation function of the tech-tetrahedron enables precise mitochondrial anchoring via the triphenylphosphine modification of a single vertex. The enzymatic control function deploys a telomerase-gated latch that opens upon primer elongation to temporarily release a self-assembly initiator. The self-assembly function uses this initiator to trigger catalytic hairpin self-assembly at two hairpin-decorated vertices, generating spatial "hand-in-hand" DNA nanoaggregates. These DNA nanoaggregates serve as polyanionic barriers that disrupt physiological traits and mitochondrial-cytoplasmic metabolite exchange, thereby impairing aerobic respiration and glycolysis, and reducing ATP production. Transcriptomic analysis revealed that the increased expression of polycystin-1 forms a feedback loop with Tech-tetrahedron, highlighting potential interference with nicotinamide nucleotide transhydrogenase activity. The resulting bioenergetic collapse led to a 24.67% increase in tumor apoptosis and a 63.16% tumor growth inhibition. Overall, Tech-tetrahedron offers a precise strategy for subcellular energy intervention and a transformative approach to targeted cellular regulation via artificial nanoaggregates.