Condensation-Enhanced DNA Nanotracker on Facilitating Spatially Selective Intracellular miRNA Profiling and Robust Gene Therapy/Chemotherapy

Intracellular miRNA imaging and cooperative gene therapy are significant for cancer diagnosis and treatment. However, the spatial-selective imaging and therapeutic potential of RNA through programmable DNA probes is extremely hindered due to the disorder, low local concentration, and easy degradation toward the sensing/therapeutic-associated DNA probes. To address this, we spatially confined sensing probes and therapeutic siRNA within a highly ordered spherical DNA condensed tracker (GREAT) engineered for activation by target miRNA-221. Compared to the DNA probes diffused in a homogeneous phase, the condensation-enhanced GREAT increased the sensitivity and the resistance to degradation under the cellular environment due to the effective ordered arrangement and crowding effect of the probes. The strand displacement reaction on GREAT condensation exhibited remarkably higher reaction efficiency compared to that of the conventional diffusive strand displacement reaction due to the effective enrichment of the DNA/RNA duplex and the spatial confinement effect, which achieve sensitive detection of miRNA-221 with a low limitation of 2.69 fM and ultraclear fluorescence imaging of intracellular miRNA-221. Specifically, under the activation of miRNA-221, the programmed GREAT condensation was able to release doxorubicin (Dox) molecules through the strand displacement reaction while simultaneously in situ generating small interfering RNA (siRNA), significantly decreasing the cellular degradability of naked siRNA and improving the therapeutic effects by integrating the chemo-therapeutic strategy. The feasibility of gene therapy by GREAT condensation was verified by the vascular endothelial growth factor (VEGF) expression in HeLa cell lines. The designed GREAT condensation provides an effective nanoplatform for sensitive biomarker detection, facilitating precise disease diagnosis and potent tumor therapy.