ATP‐fueled transient DNAzyme. a) Schematic illustration of ATP‐fueled transient DNAzyme for substrate cleavage. b) Time‐dependent AGE (2 wt.%, 105 V, 1.5 h) for transient DNAzyme. c, d) Time‐dependent FI measurements of substrate cleavage by (c) ATP‐fueled transient DNAzyme and (d) static DNAzyme. The shaded areas correspond to the standard deviation of duplicate measurements. e) Relative kinetics for RNA cleavage for both dynamic and static DNAzymes. Error bars correspond to the standard deviation of duplicate measurements. f) Time‐dependent FI measurements corresponding to multiple temporal activations of DNAzyme for controlled substrate cleavage. Conditions: b) 25 °C, 10 μM each DNA species, 0.46 WU T4 DNA ligase, 1 U BsaI, and 0.03 mM ATP. c) The same as above but added varied concentration of Substrate. d) Without BsaI, and fed with varied concentration of Substrate. f) The same as (b) but containing 2.67 μM Substrate, and repeatedly fueled with ATP.