Designer Dynamic DNA Nanoaggregate in Living Cell for Mitochondrial Energy Restriction

ABSTRACT 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.