Control of Reversible Formation and Dispersion of the Three Enzyme Networks Integrating DNA Computing

The majority of biological reactionsin the cytoplasm of livingcells occur via enzymatic cascade reactions. To achieve efficientenzyme cascade reactions mimicking the proximity conditions of enzymesin the cytoplasm, the proximity of each enzyme, creating a high localconcentration of proteins, has been recently investigated using theconjugation of synthetic polymer molecules, proteins, and nucleicacids. Although there have been methodologies reported for the complexformation and enhanced activity of cascade reactions due to the proximityof each enzyme using DNA nanotechnology, one pair of the enzyme (GOxand HRP) complex is only assembled by the mutual independence of variousshapes of the DNA structure. This study reports the network formationof three enzyme complexes assembled by a triple-branched DNA structureas a unit, thus enabling the reversible formation and dispersion ofthe three enzyme complex networks using single-stranded DNA, RNA,and enzymes. It was found that the activities of the three enzymecascade reactions in the enzyme-DNA complex network were controlledby formation and dispersion of the three enzyme complex networks,due to the proximity of each enzyme with the enzyme-DNA complexnetwork. Furthermore, three micro RNA sequences for breast cancerbiomarkers were successfully detected using an enzyme-DNA complexnetwork integrated with DNA computing. Overall, the reversible formationand dispersion of the enzyme-DNA complex network through theexternal stimulation of biomolecules and DNA computing provide a novelplatform for controlling the production amount, diagnosis, theranostics,and biological or environmental sensing.

Automated summary

The majority of biological reactions in cells occur via enzymatic cascade reactions. Researchers have recently investigated the proximity of enzymes in the cytoplasm to achieve efficient enzyme cascade reactions. They have used synthetic polymer molecules, proteins, and nucleic acids to create a high local concentration of enzymes. This study reports the reversible formation and dispersion of enzyme-DNA complex networks, controlled by the proximity of each enzyme, which provides a novel platform for controlling production, diagnosis, and biological or environmental sensing.