RADD: A real-time FRET-based biochemical assay for DNA deaminase studies.
APOBEC3
Cancer
Cytidine deaminase
Fluorescence resonance energy transfer (FRET)
High‐throughput screening (HTS)
Real-time assay
Journal
Methods in enzymology
ISSN: 1557-7988
Titre abrégé: Methods Enzymol
Pays: United States
ID NLM: 0212271
Informations de publication
Date de publication:
2024
2024
Historique:
medline:
11
10
2024
pubmed:
11
10
2024
entrez:
10
10
2024
Statut:
ppublish
Résumé
In recent years, the connection between APOBEC3 cytosine deaminases and cancer mutagenesis has become ever more apparent. This growing awareness and lack of inhibitory drugs has created a distinct need for biochemical tools that can be used to identify and characterize potential inhibitors of this family of enzymes. In response to this challenge, we have developed a Real-time APOBEC3-mediated DNA Deamination (RADD) assay. The RADD assay provides a rapid, real-time fluorescence readout of APOBEC3 DNA deamination and serves as a crucial addition to the existing APOBEC3 biochemical and cellular toolkit. This method improves upon contemporary DNA deamination assays by offering a more rapid and quantifiable readout as well as providing a platform that is readily adaptable to a high-throughput format for inhibitor discovery. In this chapter we provide a detailed guide for the usage of the RADD assay for the characterization of APOBEC3 enzymes and potential inhibitors.
Identifiants
pubmed: 39389668
pii: S0076-6879(24)00409-9
doi: 10.1016/bs.mie.2024.08.001
pii:
doi:
Substances chimiques
DNA
9007-49-2
Cytidine Deaminase
EC 3.5.4.5
Enzyme Inhibitors
0
APOBEC Deaminases
EC 3.5.4.5
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
311-345Informations de copyright
Copyright © 2024. Published by Elsevier Inc.