Nanozymes have emerged as highly promising alternatives to natural enzymes, featuring high stability, facile synthesis, tunability, and cost-effectiveness, making them key players in biocatalysis, environmental remediation and biomedical applications. However, the moderate catalytic activity and poor specificity of nanozymes hinder their broader applications. To address these limitations, researchers have focused on developing regulatory strategies that enhance the catalytic activity and specificity of nanozymes. Among them, DNA with programmability, recognition specificity, and abundant activate binding sites, greatly facilitates the development of smart nanozymes. Therefore, this review comprehensively summarizes DNA-mediated regulation strategies and recent researches progress on nanozymes in the field of biosensing. In addition, we discuss the current challenges and future perspectives regarding DNA-mediated regulation of nanozymes.