DNAzymes are widely used as functional units for creating DNA-based sensors and devices. Switching of DNAzyme activity by external stimuli is of increasing interest. Herein we report a Cu -responsive DNAzyme rationally designed by incorporating one of the most stabilizing artificial metallo-base pairs, a Cu -mediated carboxyimidazole base pair (Im -Cu -Im ), into a known RNA-cleaving DNAzyme. Cleavage of the substrate was suppressed without Cu , but the reaction proceeded efficiently in the presence of Cu ions. This is due to the induction of a catalytically active structure by Im -Cu -Im pairing. The on/off ratio was as high as 12-fold, which far exceeds that of the previously reported DNAzyme with a Cu -mediated hydroxypyridone base pair. The DNAzyme activity can be regulated specifically in response to Cu ions during the reaction through the addition, removal, or reduction of Cu . This approach should advance the development of stimuli-responsive DNA systems with a well-defined sharp switching function.
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