Highly sensitive nuclease assays based on chemically modified DNA or RNA

Area of Science:

• Biochemistry
• Molecular Biology
• Enzymology

Background:

• Nucleolytic enzymes like DNase and RNase are implicated in various diseases, including cancers and cardiovascular conditions.
• DNase I and RNase have therapeutic applications, while angiogenin, an RNase A family member, is a target for anticancer drug development.
• Accurate detection of these enzymes is crucial for disease diagnosis and therapeutic monitoring.

Purpose of the Study:

• To review fluorometric and electrochemical techniques for detecting DNase and RNase activities in disease contexts.
• To highlight the utility of these methods in disease diagnosis and drug discovery.
• To discuss the application of these techniques in analyzing biological samples like blood and serum.

Main Methods:

• Fluorometric detection using oligonucleotides with FRET-causing chromophores that become fluorescent upon enzymatic cleavage.
• Electrochemical detection employing ferrocenyl oligonucleotides, with or without ribonucleoside units, for RNase and DNase detection.
• Application of these methods to monitor disease-associated enzyme levels in blood or serum samples.

Main Results:

• FRET-based oligonucleotides offer a sensitive tool for detecting DNase and RNase activity, facilitating drug discovery.
• Electrochemical techniques provide robust detection of these nucleolytic enzymes, unaffected by common sample interferents.
• These methods have demonstrated utility in monitoring diseases linked to aberrant DNase and RNase levels.

Conclusions:

• Fluorometric and electrochemical techniques represent valuable tools for detecting DNase and RNase in disease.
• These enzyme detection methods support disease diagnosis, monitoring, and the development of novel therapeutics.
• The described techniques are adaptable for analyzing complex biological samples, enhancing their clinical relevance.