DNAzymes: Selected for Applications

TLDR

DNAzymes are single‑stranded DNA molecules with catalytic activity that, though absent in nature, can be isolated from random‑sequence pools by in‑vitro selection and have been reported to catalyze a diverse range of chemical transformations. The paper discusses newly engineered DNAzymes that mimic intriguing functions of protein enzymes, focusing on RNA‑cleaving DNAzymes whose activity depends on specific chemical or biological stimuli. The authors describe ligand‑responsive RNA‑cleaving DNAzymes used in bioanalytical assays and biosensors, including strategies that convert analyte‑activated DNAzyme activity into large amounts of DNA amplicons through rolling‑circle amplification to achieve high detection sensitivity. DNAzymes are fully compatible with DNA amplification techniques.

Abstract

Abstract DNA‐based enzymes, also known as deoxyribozymes or DNAzymes, are single‐stranded DNA molecules with catalytic activity. DNAzymes do not exist in nature but can be isolated from random‐sequence DNA pools using in vitro selection. To date, many DNAzymes that collectively catalyze a diverse range of chemical transformations have been reported. Here, examples of new DNAzymes engineered to mimic some intriguing functions of naturally occurring protein‐based enzymes are discussed. This is followed by discussions of recent examples of a particular class of DNAzymes, known as “RNA‐cleaving DNAzymes”, that have been derived specifically so that their activity is strictly dependent on a given chemical or biological stimulus. Some unique ways to employ ligand‐responsive DNAzymes for the design of bioanalytical assays and biosensors are then highlighted. Being DNA molecules, DNAzymes have proven to be entirely compatible with DNA amplification. Several approaches are then discussed, which relay the activity of an analyte‐activated DNAzyme into the production of massive amounts of DNA amplicons, via “rolling circle amplification”, in biosensing applications designed to deliver very high levels of detection sensitivity.