Electrochemical approach for the analysis of DNA degradation in native DNA and apoptotic cells

• Published in: Heliyon Vol. 10; no. 3; p. e25602
• Main Authors: Agafonova, Lyubov E., Zhdanov, Dmitry D., Gladilina, Yulia A., Shishparenok, Anastasia N., Shumyantseva, Victoria V.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 15-02-2024; Elsevier
• Subjects: Apoptosis; Bioelectrochemistry; Carbon nanotubes; DNA fragmentation; Modified electrodes; Plasmid DNA; Carbon nanotubes; Modified electrodes; Plasmid DNA; Bioelectrochemistry; DNA fragmentation; Apoptosis
• ISSN: 2405-8440; 2405-8440
• DOI: 10.1016/j.heliyon.2024.e25602

The aim of this work was to develop an electrochemical approach for the analysis of DNA degradation and fragmentation in apoptotic cells. DNA damage is considered one of the major causes of human diseases. We analyzed the cleavage processes of the circular plasmid pTagGFP2-N and calf thymus DNA, which were exposed to restriction endonucleases (the restriction endonucleases BstMC I and AluB I and the nonspecific endonuclease I). Genomic DNA from the leukemia K562 cell line was used as a marker of the early and late (mature) stages of apoptosis. Registration of direct electrochemical oxidation of nucleobases of DNA molecules subjected to restriction endonuclease or apoptosis processes was proposed for the detection of these biochemical events. Label-free differential pulse voltammetry (DPV) has been used to measure endonuclease activities and DNA damage using carbon nanotube-modified electrodes. The present DPV technique provides a promising platform for high-throughput screening of DNA hydrolases and for registering the efficiency of apoptotic processes. DPV comparative analysis of the circular plasmid pTagGFP2-N in its native supercoiled state and plasmids restricted to 4 and 23 parts revealed significant differences in their electrochemical behavior. Electrochemical analysis was fully confirmed by means of traditional methods of DNA analysis and registration of apoptotic process, such as gel electrophoresis and flow cytometry.
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•Electrochemical analysis of DNA cleavage and degradation is proposed.•DNA fragmentation was used to detect apoptotic processes.•The activity of DNA hydrolases based on DNA fragmentation was evaluated.