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bioRxiv · 10.1101/2024.05.11.593688

A real-time biochemical assay for quantitative analyses of APOBEC-catalyzed DNA deamination

Belica, C. A.·Carpenter, M. A.·Chen, Y.·Brown, W. L.·Moeller, N. H.·Boylan, I. T.·Harris, R. S.·Aihara, H.
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Abstract

Over the past decade, the connection between APOBEC3 cytosine deaminases and cancer mutagenesis has become increasingly apparent. This growing awareness has created a need for biochemical tools that can be used to identify and characterize potential inhibitors of this enzyme family. In response to this challenge, we have developed a Real-time APOBEC3-mediated DNA Deamination (RADD) assay. This assay offers a single-step set-up and real-time fluorescent read-out, and it is capable of providing insights into enzyme kinetics and also offering a high-sensitivity and easily scalable method for identifying APOBEC3 inhibitors. This assay serves as a crucial addition to the existing APOBEC3 biochemical and cellular toolkit and possesses the versatility to be readily adapted into a high-throughput format for inhibitor discovery.

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Belica, C. A., Carpenter, M. A., Chen, Y., Brown, W. L., Moeller, N. H., Boylan, I. T., Harris, R. S., Aihara, H.. 2024-05-12. A real-time biochemical assay for quantitative analyses of APOBEC-catalyzed DNA deamination. https://doi.org/10.1101/2024.05.11.593688

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