Fluorescence imaging tools enable the in situ visualization of those molecules involved in various cell signaling pathways, but directly describing these pathways instead of the separate mediators in situ is much more meaningful and still full of challenges. In this work, a dual-responsive DNA nanodevice that allows the available imaging of an apoptotic signaling pathway in living cells has been developed. The nanodevice is constructed through assembling an elaborately designed Y-shaped DNA (Y-DNA) layer on gold nanoparticles (AuNPs). Only if an apoptotic signaling pathway involving the manganese superoxide dismutase (MnSOD) mRNA and downstream cytochrome c (Cyt c) is presented to serve as the input, the nanodevice can perform an "AND" logic gate operation, disassembling the Y-DNA from the AuNP surface and thereafter outputting a fluorescence signal. In comparison with the fluorescence imaging methods that target individual specific molecules, our strategy allows direct profiling of a specific signaling pathway by connected characterization of two correlative targets. The programmable feature of this strategy also shows the potential for the profiling of other signaling pathways. The concept of studying the line connecting two points will contribute to the systematic interrogation on the signaling networks in situ as the networks are composed of lines instead of individual points.
Keywords: DNA nanodevice; cytochrome c; logic gate; mRNA; signaling pathway.