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In vivo cII, gpt, and Spi⁻ gene mutation assays in transgenic mice and rats

Mugimane G Manjanatha1, Xuefei Cao, Sharon D Shelton

  • 1Division of Genetic and Molecular Toxicology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 31, 2013
PubMed

Insights

This study details methods for transgenic mutation assays using Big Blue®, Muta™ mouse, and gpt-delta models. These assays identify genotoxic hazards and carcinogen mechanisms by analyzing reporter gene mutations in rodents.

Area of Science:

  • Toxicology and Genetic Toxicology
  • Molecular Biology and Carcinogenesis

Background:

  • Transgenic mutation assays are crucial for identifying genotoxic hazards and understanding carcinogen mechanisms.
  • Popular models include Big Blue® (rat/mouse), Muta™ mouse, and gpt-delta (rat/mouse), each utilizing distinct reporter genes.

Purpose of the Study:

  • To describe standardized methodologies for conducting mutation assays in established transgenic rodent models.
  • To provide a framework for the recovery, analysis, and quantification of mutations in reporter genes.

Main Methods:

  • Isolation of transgenes from tissue DNA.
  • Packaging of transgenes into lambda phage for bacterial infection.
  • Selective isolation and quantification of mutant plaques (cII, Spi⁻) and colonies (gpt).

Main Results:

  • Detailed procedures for implementing mutation assays in Big Blue®, Muta™ mouse, and gpt-delta systems.
  • Methodology allows for the quantification of mutations within specific reporter genes (cII, gpt) and loci (Spi⁻).

Conclusions:

  • The described methodologies enable robust characterization of genotoxic effects and carcinogen modes of action.
  • Further analysis of selected mutants can identify specific sequence alterations and large deletions.

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