Intelligent Reconfiguration‐Promoted Cellular Internalization of Core–Shell DNA Nanoprobe Equipped with Successive Dual Stimuli‐Responsive Protective Satellites for Amplification Fluorescence Imaging of Tumor Cells

Abstract Although DNA probes have attracted increasing interest for precise tumor cell identification by imaging intracellular biomarkers, the requirement of commercial transfection reagents, limited targeting ligands, and/or non‐biocompatible inorganic nanostructures has hampered the clinic translation. To circumvent these shortcomings, a reconfigurable ES‐NC (Na + ‐dependent DNAzyme (E)‐based substrate (S) cleavage core/shell DNA nanocluster (NC)) entirely from DNA strands is assembled for precise imaging of cancerous cells in a successive dual‐stimuli‐responsive manner. This nanoprobe is composed of a strung DNA tetrahedral satellites‐based protective (DTP) shell, parallelly aligned target‐responsive sensing (PTS) interlayer, and hydrophobic cholesterol‐packed innermost layer (HCI core). Tetrahedral axial rotation‐activated reconfiguration of DTP shell promotes the exposure of interior hydrophobic moieties, enabling cholesterol‐mediated cellular internalization without auxiliary elements. Within cells, over‐expressed glutathione triggers the disassembly of the DTP protective shell (first stimulus), facilitating target‐stimulated signal transduction/amplification process (second stimuli). Target miRNA‐21 is detected down to 10.6 fM without interference from coexisting miRNAs. Compared with transfection reagent‐mediated counterpart, ES‐NC displays a higher imaging ability, resists nuclease degradation, and has no detectable damage to healthy cells. The blind test demonstrates that the ES‐NC is suitable for the identification of cancerous cells from healthy cells, indicating a promising tool for early diagnosis and prediction of cancer.