Precise Synergistic Photoregulation of Dual‐Target Gene Expression by Engineering Caged Two‐in‐One DNA‐RNA Nanoframework Based on Near‐Infrared Light Uncaging Strategy

The synergistic regulation of gene expression at spatiotemporal resolution is of great importance for exploring specific gene functions and regulating biological processes. Here, we have designed and developed an engineering caged two-in-one DNA-RNA nanoframework woven with near-infrared responsive divalent siRNAs, which can be used for the synergistic photoregulation of dual-target tumor gene expression. In-situ generated reactive oxygen species using near-infrared light can release dual-target divalent siRNAs from caged DNA-RNA tetrahedron through the scission reaction induced by reactive oxygen species. The caged two-in-one DNA-RNA nanoframework was temporarily inhibited from RNAi-induced gene silencing activity without light irradiation. Near-infrared light irradiation effectively blocked the synergistic driving effects of two key cancer oncogenes, CENPF and FOXM1, through precise photoregulation of dual-target gene expression and tumorigenic PI3K and MAPK signaling pathways to achieve the cellular synergistic anti-tumor effects of combinatorial RNAi. The self-assembled caged two-in-one DNA-RNA nanoframework achieved the simultaneous photoregulation of dual-target RNAi-induced gene silencing in living tumor cells with spatiotemporal resolution. Furthermore, the NIR-responsive antitumor effect of the caged two-in-one DNA-RNA nanoframework was also achieved in vivo. This engineering caged two-in-one DNA-RNA nanoframework will provide a promising toolbox and strategy for exploring gene regulatory networks and divalent siRNA-based precise therapies.