Spatiotemporally Controlled Protein Corona‐Armored DNA Nanomachine for Multitype Biomarker Imaging in Living Cells

Abstract DNA nanomachines have significant potential for biosensing because of their high programmability, signal amplification ability, and excellent biocompatibility. However, under physiological conditions, their performance is often hindered by insufficient endogenous driving forces, nuclease degradation, and lysosomal entrapment. In this study, we develop a protein corona (PC)‐armored DNA nanomachine with spatiotemporal control for multiple intracellular biomarker imaging. By pre‐assembling a UV‐responsive PC shell, the DNA nanomachine forms a physical barrier that protects nucleic acids from nucleases, reduces uncontrollable PC formation during unimpaired operation, and enhances cellular uptake via PC‐mediated targeting. After internalization, the retained PC promotes lysosomal escape, followed by light‐triggered disassembly for precise cytoplasmic operation of the nanomachine. Notably, our nanomachine employs RNA as a track and the widely expressed intracellular RNase H enzyme as the driving force, circumventing sequence limitations and establishing a general operational platform. Through this modulation, we achieve in situ imaging of microRNA in U87 human glioblastoma cells and ATP in A549 lung carcinoma cells, pioneering a PC‐enabled paradigm for DNA nanomachines and building a versatile intracellular diagnostic platform for precision medicine.