Phenoxy radical reactivity of nucleic acids: practical implications for biotinylation
Name
cbic.202000854.pdf
Description
Published version
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754.16 KB
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Author(s)
Wilbanks, Brandon
Garcia, Brian
Byrne, Shane
Dedon, Peter C
Maher, L James
Date Issued
2021
Journal
ChemBioChem
Publisher
Wiley
Version
Final published version
Abstract
Recent advances in peroxidase-mediated biotin tyramide (BT) signal amplification technology have resulted in high-resolution and subcellular compartment-specific mapping of protein and RNA localization. Horseradish peroxidase (HRP) in the presence of H O is known to activate phenolic compounds for phenoxy radical reaction with nucleic acids, where biotinylation by BT is a practical example. BT reactivity with RNA and DNA is not understood in detail. We report that BT phenoxy radicals react in a sequence-independent manner with guanosine bases in RNA. In contrast, DNA reactivity with BT cannot be detected by our methods under the same conditions. Remarkably, we show that fluorescein conjugates DNA rapidly and selectively reacts with BT phenoxy radicals, allowing convenient and practical biotinylation of DNA on fluorescein with retention of fluorescence. 2 2
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1002/CBIC.202000854
