Bifunctional fluorescent aptasensors for simultaneously monitoring multiple signaling molecules involved in cellular...

ATP and SO₂, as two crucial signaling molecules, are extensively involved in various physiological and pathological processes in organisms, and also play important roles in cellular inflammation. Detecting and imaging the secreted ATP and SO₂ from inflammatory cells can help monitor the inflammatory process and dynamically evaluate the effects of drug treatment. To this end, here we design a DNA nanotweezer-based aptasensor incorporating two i-motif sequences that can selectively bind to a fluorescent ligand and a SO₂-responsive fluorescent probe, which generate fluorescence signals in response of ATP and SO₂, respectively. It enables the development of a cell-surface-anchored bifunctional fluorescent DNA sensor for simultaneously monitoring the release of endogenous ATP and SO₂ during drug-induced cellular inflammation process, revealing that these two molecules can serve as specific signaling molecules involved in cellular inflammation, distinguishing from other pathological responses. This highlights the great potential of our designed aptasensor for dynamically evaluating cellular physiological and pathological processes.