This work reports exquisite engineering of catalytic activity of DNA‐templated silver nanoclusters (DNA‐AgNCs) based on unique adsorption phenomena of DNAs on DNA‐AgNCs and reversible transition between double and triple‐stranded DNAs. Four DNA homopolymers exhibit different inhibition effects on the catalytic activity of DNA‐AgNCs, poly adenine (polyA) > poly guanine (polyG) > poly cytosine (polyC) > poly thymine (polyT), demonstrating that polyA strands have the strongest adsorption affinity on DNA‐AgNCs. Through the formation of T‐A•T triplex DNAs, catalytic activity of DNA‐AgNCs is restored from the deactivated state by double or single‐stranded DNAs, indicating the participation of N7 groups of adenine bases in binding to DNA‐AgNCs and blocking active sites. Accordingly, reversibly regulating catalytic activity of DNA‐AgNCs can be realized based on DNA input‐stimulated transition between duplex and triplex structures. In the end, two low‐cost and facile biosensing methods are presented, which are derived from the activity‐switchable platform. It is worthy to anticipate that the DNA‐AgNCs with controlled catalytic activity will inspire researchers to devise more functionalized nanocatalysts and contribute to the exploration of intelligent biomedicine in the future.