Methods to Assess Oxidative DNA Base Damage Repair of Apurinic/Apyrimidinic (AP) Sites Using Radioactive and Nonradioactive Oligonucleotide-Based Assays

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dc.contributor.authorGupta, Kunj Bihari
dc.contributor.authorKaur, Sharanjot
dc.contributor.authorDhiman, Monisha
dc.contributor.authorMantha, Anil Kumar
dc.date.accessioned2024-01-21T10:37:03Z
dc.date.accessioned2024-08-13T11:19:34Z
dc.date.available2024-01-21T10:37:03Z
dc.date.available2024-08-13T11:19:34Z
dc.date.issued2022-01-19T00:00:00
dc.description.abstractReactive oxygen species (ROS) overproduction results in oxidative stress leading to genomic instability via the generation of small base lesions in the genome, and this unrepaired DNA base damage leads to various cellular consequences. The oxidative stress-mediated DNA base damage is involved in various human disorders like cancer, cardiovascular, ocular, and neurodegenerative diseases. Base excision repair (BER) pathway, one of the DNA repair pathways, is majorly involved in the repair of oxidative DNA base lesions, which utilizes a different set of enzymes, including endonuclease viz Apurinic/apyrimidinic endonuclease 1 (APE1). APE1 is a well-known multifunctional enzyme with DNA repair, REDOX regulatory, and protein-protein interaction/cross-talk functions associated with the cell survival mechanisms. APE1 acts as an important player in both normal and cancerous cell survival; thus, evaluating its endonuclease activity in the biological samples provide useful readout of the DNA repair capacity/ability, which can be used to tune for the development of therapeutic candidates via either stimulating or blocking its DNA repair function in normal vs. cancer cells, respectively. This chapter enlists two methods used for the determination of APE1's endonuclease activity by oligonucleotide-based radioactive P32-labeled and nonradioactive fluorescence dyes using the cell extracts and recombinant APE1 protein. � 2022. Springer Science+Business Media, LLC, part of Springer Nature.en_US
dc.identifier.doi10.1007/978-1-0716-1896-7_16
dc.identifier.issn19406029
dc.identifier.urihttp://10.2.3.109/handle/32116/3468
dc.identifier.urlhttps://link.springer.com/10.1007/978-1-0716-1896-7_16
dc.language.isoen_USen_US
dc.publisherNLM (Medline)en_US
dc.subjectAP siteen_US
dc.subjectAPE1 activityen_US
dc.subjectApurinic/Apyrimidinic endonuclease 1 (APE1)en_US
dc.subjectDenaturing PAGEen_US
dc.subjectFluoresceinen_US
dc.titleMethods to Assess Oxidative DNA Base Damage Repair of Apurinic/Apyrimidinic (AP) Sites Using Radioactive and Nonradioactive Oligonucleotide-Based Assaysen_US
dc.title.journalMethods in molecular biology (Clifton, N.J.)en_US
dc.typeArticleen_US
dc.type.accesstypeClosed Accessen_US

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