Silver is a very common heavy metal, and its detection is of significant analytical importance.DNAzymes are DNA-based catalysts; they typically recruit divalent and trivalent metal ions for catalysis. Herein, we report a silver-specific RNA-cleaving DNAzyme named Ag10c obtained after six rounds of in vitro selection. Ag10c displays a catalytic rate of 0.41 min -1 with 10 µM Ag + at pH 7.5 with 200 mM NaNO3, while its activity is completely inhibited with the same concentration of NaCl. Ag10c is highly specific for Ag + among all the tested metals. A catalytic beacon biosensor is designed by labeling a fluorophore and a quencher on the DNAzyme.Fluorescence enhancement is observed in the presence of Ag + with a detection limit of 24.9 nM Ag + . The sensor shows a similar analytical performance in Lake Huron water. This is the first monovalent transition metal dependent RNA-cleaving DNAzyme. Apart from its biosensor application, this study strengthens the idea of exploring beyond the traditional understanding of multivalent ion dependent DNAzyme catalysis.3