Work

Functional DNA Repair Profiling in Translational Medicine: Benchmarking Comet, γH2AX, and NGS Assays Against Clinical Constraints

Year: 2026

Type: article

Abstract: Quantifying DNA repair capacity (DRC) is pivotal for stratifying patients in oncology and autoimmune disorders, yet methodological heterogeneity compromises data reproducibility. While basic research relies on genetically encoded reporters, translational settings demand robust assays compatible with biobanked material, particularly Peripheral Blood Mononuclear Cells (PBMCs). This review benchmarks functional DNA repair assays-ranging from alkaline/neutral comet variants and high-content foci imaging (γH2AX/53BP1) to emerging Next-generation sequencing (NGS)-based break mapping-against the rigors of clinical application. We critically evaluate sensitivity, specificity, and throughput, identifying artifacts introduced by cryopreservation, steroid therapy, and oxidative stress. Furthermore, we propose a "Minimum Reporting Standard" checklist to harmonize DRC quantification. By distinguishing established validation tools from experimental artifacts, this framework aligns assay selection with specific biological endpoints and clinical feasibility.

Source: Current Issues in Molecular Biology

DOI: 10.3390/cimb48020149

Authors: Anna Macieja, Marta Poplawska, Karolina Przybylowska-Sygut, Joanna Makowska, Tomasz Poplawski

Institution: Medical University of Lodz

Language: English

Topic: DNA Repair Mechanisms

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $2,654