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Related Experiment Videos

A model for testing recombinogenic sequences in the mouse germline

M E Moynahan1, E Akgün, M Jasin

  • 1Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.

Human Molecular Genetics
|July 1, 1996
PubMed
Summary

Researchers developed a transgenic mouse model to study genetic recombination in male germ cells. This system quantifies recombination events, aiding research into human genetics and genomic instability.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Reproductive Biology

Background:

  • Homologous recombination is crucial for species diversity and gene evolution.
  • Mammalian germline recombination studies, particularly for human genetics, lack well-developed systems.
  • Understanding germline recombination is key to investigating genomic instability.

Purpose of the Study:

  • To establish a transgenic mouse system for studying recombination in the male germline.
  • To develop a quantifiable method for detecting recombination events in sperm.
  • To investigate recombination within specific human DNA sequences implicated in genetic disease.

Main Methods:

  • Utilized an intron-interrupted lacZ reporter gene in a transgenic mouse model.
  • Employed flow cytometry with fluorogenic substrates to detect beta-galactosidase positive sperm.

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  • Confirmed germline recombination through histochemical staining, mRNA analysis, and PCR of sorted sperm.

    • Main Results:

    • Quantified recombination within a 1.7 kb human glycophorin breakpoint cluster region repeat.
    • Detected approximately 0.04-0.09% fluorescent sperm, indicating successful recombination.
    • Validated the findings through multiple molecular and cellular analyses.

    Conclusions:

    • The developed transgenic mouse system effectively studies male germline recombination.
    • This system is adaptable for analyzing sequences associated with elevated recombination and human genetic diseases.
    • Further research can provide insights into chromosomal instability mechanisms.