Qualitative and Quantitative Analysis of DNA Cytidine Deaminase Activity.
Methods Mol Biol
; 2444: 161-169, 2022.
Article
em En
| MEDLINE
| ID: mdl-35290637
ABSTRACT
The human genome encodes eleven DNA cytidine deaminases in the AID/APOBEC family, which encompass endogenous roles ranging from genetic diversification of the immunoglobulin locus to virus restriction. All AID/APOBEC functions are enabled by their catalyzation of cytidine deamination resulting in mutations and DNA damage. When acting aberrantly, deaminases can cause off-target mutations in the cellular genome resulting in somatic mutations, DNA damage, and genome instability. An association between cytidine deaminase-induced mutations and human cancers has been recognized over the last decade, necessitating assays for investigation of intracellular deaminase activity. Here we present two assays for deamination activity which enable in vitro evaluation of in vivo events. We define both a qualitative assay to confirm deaminase activity within cells as well as a quantitative assay for granular evaluation and comparisons of deamination activity across different cell populations or experimental conditions. The two procedures are customizable assays which can easily be adapted to individual labs and experiments.
Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
DNA
/
Citidina Desaminase
Tipo de estudo:
Qualitative_research
Limite:
Humans
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
Estados Unidos