Live-cell imaging of adenosine triphosphate (ATP) is crucial for gaining insight into enzymatic activities and biological processes. However, it is still difficult to achieve sensitive and non-destructive ATP imaging due to the difficulty of amplifying molecules within cells and the complexity of intercellular environments. Herein, we present non-enzymatic ATP aptamer-based DNAzyme (Aptazyme)-driven autonomous DNA circuits for sensitive ATP imaging in living cells. The combination of dual amplifiers with aptazyme and catalysis hairpin assembly (CHA) circuits yielded a detection limit of 0.21 μM for ATP, which was 83.9 times more sensitive than the aptazyme-based probe without CHA circuits amplification. Moreover, the efficient carrier function and quenching effect of molybdenum disulfide (MoS2) made it possible to incorporate circuits components into cells without the requirement of transfection, thus allowing for efficient and non-destructive ATP detection within live cells. Therefore, the non-enzymatic aptazyme-driven autonomous DNA circuits present a promising novel approach for the sensitive detection of small molecules within live cells.