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Updated: May 26, 2026

Identifying DNA Mutations in Purified Hematopoietic Stem/Progenitor Cells
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Mammalian cell HPRT gene mutation assay: test methods.

George E Johnson1

  • 1DNA Damage Group, School of Medicine, Swansea University, Swansea, Wales, UK. G.Johnson@swansea.ac.uk

Methods in Molecular Biology (Clifton, N.J.)
|December 8, 2011
PubMed
Summary

The hypoxanthine phosphoribosyl transferase (HPRT) gene mutation assay detects a broad range of mutagens in mammalian cells. This assay is crucial for identifying mammalian-specific mutagenic agents missed by other tests.

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Area of Science:

  • Toxicology
  • Genetics
  • Molecular Biology

Background:

  • Standard mutagenicity testing may miss mammalian-specific agents.
  • The hypoxanthine phosphoribosyl transferase (HPRT) gene on the X chromosome is a sensitive target for mutation detection.
  • Existing guidelines recommend a third assay for comprehensive mutagenicity testing.

Purpose of the Study:

  • To detail the methodology of the HPRT gene mutation assay in mammalian cells.
  • To highlight the assay's utility in detecting a wide spectrum of mutagens.
  • To provide technical guidance for conducting the HPRT assay.

Main Methods:

  • Utilizes the hypoxanthine phosphoribosyl transferase (HPRT) gene as a model in mammalian cell lines.
  • Employs positive selection with a toxic analogue to identify HPRT-deficient mutants.

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  • Methodology is similar to the thymidine kinase (TK) mouse lymphoma assay (MLA).

    • Main Results:

    • The HPRT assay detects mutations that abolish gene or protein function, resulting in viable mutant colonies.
    • It responds to a broad range of mutagens, including those causing DNA damage.
    • Human cells are readily applicable, and modifications allow for mechanistic studies.

    Conclusions:

    • The HPRT assay is a valuable tool for identifying mammalian-specific mutagenic agents.
    • It complements bacterial gene mutation assays and chromosomal aberration tests.
    • The assay provides insights into the mode of action of chemical compounds.