Silver nanoclusters are a class of fluorophores with excellent physical and chemical properties and have emerged as an attractive tool in biological applications. Using nucleic acid as a template to synthesize silver nanoclusters improves their stability and endows more interesting properties. Here, we aimed to provide a comprehensive review of the advances in nucleic acid-mediated silver nanoclusters that first emerged in 2007. Focus was placed on an overview of the properties and structures of nucleic acid-templated silver nanoclusters, followed by a discussion of the regulation of the properties of silver nanoclusters by nucleic acid sequences, conformation, and structure. Subsequently, the most recent advances in nucleic acid-templated silver nanoclusters in biological detection are highlighted. Finally, current challenges, prospects, and future directions are envisioned on nucleic acid-templated silver nanoclusters.