Selection of a metal ligand modified DNAzyme for detecting Ni²

Nickel is a highly important metal, and the detection of Ni²⁺ using biosensors is a long-stand analytical challenge. DNA has been widely used for metal detection, although no DNA-based sensors were reported for Ni²⁺. DNAzymes are DNA-based catalysts, and they recruit metal ions for catalysis. In this work, in vitro selection of RNA-cleaving DNAzymes was carried out using a library containing a region of 50 random nucleotides in the presence of Ni²⁺. To increase Ni²⁺ binding, a glycyl-histidine-functionalized tertiary amine moiety was inserted at the cleavage junction. A representative DNAzyme named Ni03 showed a high cleavage yield with Ni²⁺ and it was further studied. After truncation, the optimal sequence of Ni03l could bind one Ni²⁺ or two Co²⁺ for catalysis, while other metal ions were inactive. Its cleavage rates for 100 μM Ni²⁺ reached 0.63 h^-1 at pH 8.0. A catalytic beacon biosensor was designed by labeling a fluorophore and a quencher on the Ni03l DNAzyme. Fluorescence enhancement was observed in the presence of Ni²⁺ with a detection limit of 12.9 μM. The sensor was also tested in spiked Lake Ontario water achieving a similar sensitivity. This is another example of using single-site modified DNAzyme for sensing transition metal ions.