Standards for Quantitative Measurement of DNA Damage in Mammalian Cells

Abstract

As the potential applications of DNA diagnostics continue to expand, there is a need for improved methods and standards for DNA analysis. This report describes several methods that could be considered for the production of reference materials for the quantitative measurement of DNA damage in mammalian cells. With the focus on DNA strand breaks, potentially useful methods for assessing DNA damage in mammalian cells are reviewed. The advantages and limitations of each method, as well as additional concerns with respect to reference material development, are also discussed. In conclusion, we outline strategies for developing candidate DNA damage reference materials that could be adopted by research laboratories in a wide variety of applications.

Cosmic Whirl: Navigating the Comet Trail in DNA: H2AX Phosphorylation and the Enigma of Uncertain Significance Variants

“…According to standards for the quantitative measurement of DNA damage, DNA DSBs created by anti-tumor drugs could be measured by comet and γH2AX assays [ 16 ]. Firstly, we used the alkaline comet assay to measure basal and Dox-induced DNA damage.…”

“…Multiple genetic and biochemical functional tests have been published, demonstrating BRCA2’s role in DNA damage by regulating the homologous recombination (HR) repair pathway and interacting with DNA repair proteins such as phosphorylated histone H2AX (γH2AX) and RAD51 [ 14 , 15 ]. According to standards for the quantitative measurement of DNA damage, DNA double-strand breaks (DSBs) created by anti-tumor drugs are measured by single-cell alkaline gel electrophoresis (comet) and γH2AX protein expression [ 16 ]. Doxorubicin (Dox), a potent anthracycline antibiotic, has multiple anticancer activities, including DSBs, reactive oxygen species (ROS) production, apoptosis, senescence, autophagy, ferroptosis, and pyroptosis, as well as the ability to modulate the immune system [ 17 ].…”