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Description

Dioxyribonuclease I (DNase I, DC 3.1.21.1) is an enzyme that cleaves DNA at phosphodiester linkages. The deviation of DNase I activity is a significant important marker for several diseases. For example, the lower activity of DNase can cause arthritis and several other autoimmune diseases, whereas higher DNase I activities are observed in patients with breast and oral cavity cancers. This makes monitoring the DNase I activity important for clinical and point-of-care diagnostics.Several methods were developed to assay DNase I activity, such as the enzyme-linked immunosorbent assay (ELISA), fluorimetric assays, radial immunodiffusion (RID) and radial enzyme diffusion (RED) assays. These methods are relatively expensive, time-consuming, and require trained personnel and laboratory equipment. Electrochemical biosensors were developed to detect DNase I activity. For this, the electrode is modified with a DNA layer and incubated in a solution containing a redox label non-covalently binding to the DNA. Then, the electrode is transferred to the redox label-free solution, and the amount of adsorbed redox label is measured by voltammetric or coulometric methods. After this, the electrode is exposed to the sample, and the measurement is repeated. As a result, the difference between the initial and final signals allows for calculating the loss of the DNA on the electrode surface proportional to the DNase I activity. This procedure has several drawbacks, such as many experimental steps and the desorption of the loss of a non-covalently bound redox label before the signal is measured.In this work, we developed an electrochemical DNA biosensor utilizing [Ru(NH3)6]3+ as a redox label that overcomes the described limitations. The signal is measured before and after the exposure to the sample in the redox label containing the solution preventing the loss of adsorbed redox label. This biosensor operates in coulometric and voltammetric regimes and is easy to use and sensitive. Importantly, this biosensor has the potential to be used in combination with the newly discovered CRISPR Cas-nucleases for the detection of DNA. The details will be discussed.

Publication Date

12-5-2022

City

Logan, UT

Keywords

autoimmune diseases, biosensor, DNase I, redox label

Disciplines

Life Sciences

Electrochemical DNA-Based Biosensor for the Detection of DNase I Activity for Potential Application for CRISPR-Cas Based Biosensor

Included in

Life Sciences Commons

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