Despite their importance, the activities of numerous human DNA nucleases are still debated (e.g. All of these can result from aberrations in the structural and/or catalytic functions of DNA and RNA nucleases (reviewed in 1, 2). The complex roles DNA and RNA nucleases play in DNA repair pathways underpin several premature ageing-, immune-, and tumour- related syndromes. They dictate many DNA repair pathway choices by controlling the DNA damage substrates created that dictate downstream options during the signalling cascade. These nucleases are involved in DNA repair, immune regulation, and have been associated with various diseases, including cancer and immune disorders.ĭNA and RNA nucleases are a hallmark of a growing number of cell signalling cascades including the DNA damage response and immune diversification. We have also extended our analysis to study the kinetics of human single-strand DNA nuclease TREX2, DNA polymerases, RNA, and RNA:DNA nucleases. The assay is sensitive enough to detect kinetics of repair enzymes when confronted with DNA mismatches or DNA methylation sites. In conjunction with a substrate library we can now analyse nuclease catalytic rates, directionality, and substrate preferences.
Using fluorescence-based methods, we present a quick, safe, cost-effective, and real-time versatile nuclease assay, which uniquely studies nuclease enzyme kinetics. Elucidating nuclease substrate specificities and co-factors can support a more definitive understanding of cellular mechanisms in physiology and disease.
Nevertheless, many nucleases have unknown or poorly characterized activities. DNA and RNA nucleases play a critical role in a growing number of cellular processes ranging from DNA repair to immune surveillance.
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