Abstract
Nucleolytic enzymes are associated with various diseases, and several methods have been developed for their detection. DNase expression is modulated in such diseases as acute myocardial infarction, transient myocardial ischemia, oral cancer, stomach cancer, and malignant lymphoma, and DNase I is used in cystic fibroma therapy. RNase is used to treat mesothelial cancer because of its antiproliferative, cytotoxic, and antineoplastic activities. Angiogenin, an angiogenic factor, is a member of the RNase A family. Angiogenin inhibitors are being developed as anticancer drugs. In this review, we describe fluorometric and electrochemical techniques for detecting DNase and RNase in disease. Oligonucleotides having fluorescence resonance energy transfer (FRET)-causing chromophores are non-fluorescent by themselves, yet become fluorescent upon cleavage by DNase or RNase. These oligonucleotides serve as a powerful tool to detect activities of these enzymes and provide a basis for drug discovery. In electrochemical techniques, ferrocenyl oligonucleotides with or without a ribonucleoside unit are used for the detection of RNase or DNase. This technique has been used to monitor blood or serum samples in several diseases associated with DNase and RNase and is unaffected by interferents in these sample types.
Highlights
Nucleases cleave or digest the phosphodiester bonds of DNA [1], RNA [2], and/or DNA-RNA hybrids [3]
DNase I is required to remove DNA from samples used in mRNA expression assays, whereas RNase A is used to remove RNA from samples used for DNA
When the rC of FcODN on the electrode was cleaved by RNase A, FcODN fragments containing ferrocene were released, reducing the current, which is detected as RNase A activity
Summary
Nucleases cleave or digest the phosphodiester bonds of DNA [1], RNA [2], and/or DNA-RNA hybrids [3]. Nucleases prevent invasion or infection by viruses carrying single- or double-stranded nucleic acids, and they are expressed in the saliva, sudoriferous glands, and skin [1,2,3]. These enzymes are called deoxyribonuclease (DNase) or ribonuclease (RNase) for their respective substrates [1,2,4]. Immunochemical detection using 794 bp PCR product labeled fluorescein and biotin
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