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Related Concept Videos

Nucleotide Excision Repair01:38

Nucleotide Excision Repair

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DNA Distortion and Damage
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
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Related Experiment Video

Updated: Oct 5, 2025

Preparation of Peripheral Blood Mononuclear Cell Pellets and Plasma from a Single Blood Draw at Clinical Trial Sites for Biomarker Analysis
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DNA Repair Capacity for Personalizing Risk and Treatment Response - Assay Development and Optimization in Human

Nawar Al Nasrallah1, Huaxin Zhou2, Patricia A Smith2

  • 1Indiana University School of Medicine, Indianapolis, IN, USA; Richard L. Roudebush VA Medical Center, Indianapolis, IN, USA.

DNA Repair
|January 27, 2022
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Summary

We developed a flow cytometry assay to measure DNA repair capacity (DRC) in cells. This assay can help personalize cancer treatment by assessing individual DNA repair abilities.

Keywords:
Lung cancerNon-homologous end-joiningNucleotide excision repair

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cancer Research

Background:

  • DNA repair capacity (DRC) is crucial for cellular health and disease, particularly in cancers.
  • Differential DRC influences disease progression and treatment response.
  • Accurate measurement of DRC is needed for translational research and personalized medicine.

Purpose of the Study:

  • To develop and optimize a flow cytometry-based assay for measuring individual DNA repair capacity.
  • To assess the utility of this assay for both nucleotide excision repair (NER) and nonhomologous end joining (NHEJ) pathways.
  • To evaluate the potential of DRC measurement in lung cancer patients.

Main Methods:

  • Developed a flow cytometry assay using GFP-expressing plasmids for NER and NHEJ.
  • Utilized ultraviolet light for NER induction and restriction enzymes for NHEJ induction.
  • Validated assay specificity using Ku80-/- and XPC-deficient cells.
  • Tested assay reproducibility and sensitivity with cell mixing assays and drug treatments (NU7441).
  • Applied the assay to peripheral blood mononuclear cells (PBMCs) from healthy volunteers and lung cancer patients.

Main Results:

  • The assay demonstrated pathway specificity for both NER and NHEJ.
  • NHEJ DRC showed a linear correlation with Ku80 concentration and decreased with NU7441 treatment.
  • NER DRC showed a linear correlation with XPC concentration.
  • Assays exhibited low interindividual/inter-assay variability and were quantifiable and reproducible in PBMCs.
  • Lung cancer patients showed different DRC measurements compared to healthy volunteers.

Conclusions:

  • The developed flow cytometry assay reliably measures individual NER and NHEJ DNA repair capacity in human PBMCs.
  • This assay is quantifiable, reproducible, and sensitive to pathway inhibition.
  • DRC measurements may offer insights into personalized disease risk and treatment strategies for cancers.
  • Further investigation is warranted to integrate DRC assessment into clinical practice for lung cancer patients.