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Related Concept Videos

DNA Damage Can Stall the Cell Cycle02:36

DNA Damage Can Stall the Cell Cycle

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In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
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In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
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Overview of DNA Repair02:25

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In order to be passed through generations, genomic DNA must be undamaged and error-free. However, every day, DNA in a cell undergoes several thousand to a million damaging events by natural causes and external factors. Ionizing radiation such as UV rays, free radicals produced during cellular respiration, and hydrolytic damage from metabolic reactions can alter the structure of DNA. Damages caused include single-base alteration, base dimerization, chain breaks, and cross-linkage.
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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
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Translesion DNA Polymerases02:10

Translesion DNA Polymerases

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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Sustainable Development

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As the human population continues to grow and use resources, we must be mindful of our planet’s natural limits. Sustainable development provides a pathway to maintain and improve human life now while also ensuring that future generations will have the resources that they need. The long-term success of sustainability efforts rests on understanding the interplay between human actions and ecological systems.
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Related Experiment Video

Updated: Jan 29, 2026

Assaying DNA Damage in Hippocampal Neurons Using the Comet Assay
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Development of a DNA damage assay system using stable human hepatocytes.

Masayuki Mishima1, Kazuki Izawa2, Masataka Tsuda2

  • 1Division of Genome Safety Science, National Institute of Health Sciences (NIHS), 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki, Kanagawa, 210-9501, Japan. mishimamsy@nihs.go.jp.

Genes and Environment : the Official Journal of the Japanese Environmental Mutagen Society
|January 28, 2026
PubMed
Summary

A new assay using stable human hepatocytes detects DNA damage, overcoming species differences in toxicology. This method provides a reliable tool for genotoxicity testing that better reflects human metabolism.

Keywords:
GenotoxicityHepaSH cellsHuman hepatocytesLiver s9Metabolic activationγh2AX

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Area of Science:

  • Toxicology
  • Genetics
  • Biochemistry

Background:

  • Species differences in metabolism pose challenges for toxicological studies.
  • Human metabolic systems are highly variable, limiting standardized in vitro genotoxicity testing.
  • There is a need for genotoxicity assays that better mimic human physiology.

Purpose of the Study:

  • To develop and validate an in-cell ELISA system for measuring DNA damage in stable human hepatocytes.
  • To assess the utility of the γH2AX marker in human hepatocytes for genotoxicity evaluation.
  • To establish optimal assay conditions for detecting genotoxic responses in a human-relevant system.

Main Methods:

  • Development of an in-cell ELISA measuring γH2AX (DNA damage marker) in stable human hepatocytes (HepaSH cells).
  • HepaSH cells stably express metabolic enzymes and drug transporters, mimicking human physiology.
  • Exposure of HepaSH cells to known indirect mutagens (e.g., benzo(a)pyrene) to assess dose-dependent DNA damage.

Main Results:

  • The developed γH2AX-SHE assay successfully detected dose-dependent increases in DNA damage in response to indirect mutagens.
  • A treatment duration of 16 hours or longer was required for genotoxic response detection.
  • High cytotoxicity (48-hour exposure) could interfere with accurate γH2AX quantification.

Conclusions:

  • The γH2AX-SHE assay is a valuable tool for detecting DNA damage in a human-relevant metabolic context.
  • Recommended assay conditions include a 24-hour treatment period and DMSO concentration ≤1%.
  • Responses at highly cytotoxic doses (<60% cell survival) require careful interpretation due to potential lack of biological relevance.