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Chromosomal insertion of foreign DNA

J O Bishop1

  • 1Centre of Genome Research, University of Edinburgh, UK.

Reproduction, Nutrition, Development
|January 1, 1996
PubMed
Summary

Generating transgenic animals relies on microinjecting DNA into embryos. This DNA forms extrachromosomal arrays, integrating into chromosomes via illegitimate recombination, often causing expression issues and mosaicism in founder animals.

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Area of Science:

  • Transgenic animal generation
  • Molecular biology
  • Genetics

Background:

  • Microinjection of DNA into early embryos is the primary method for creating transgenic animals.
  • Introduced DNA typically forms extrachromosomal concatemers through homologous recombination.
  • This process involves circularization and random cleavage of DNA molecules.

Purpose of the Study:

  • To elucidate the mechanisms of DNA concatemer formation and chromosomal integration in transgenic animals.
  • To understand the origins of mosaicism and chromosomal position effects impacting gene expression.
  • To explore strategies for improving the efficiency of generating transgenic founders with desired expression levels.

Main Methods:

  • Analysis of DNA recombination processes in microinjected embryos and cultured mammalian cells.
  • Investigation of extrachromosomal concatemer formation and subsequent chromosomal integration.
  • Evaluation of the impact of integration site on foreign gene expression.

Main Results:

  • Evidence suggests homologous recombination generates extrachromosomal DNA arrays.
  • Chromosomal integration occurs randomly via illegitimate recombination, often during DNA replication.
  • This integration process frequently results in mosaic transgenic animals and chromosomal position effects.

Conclusions:

  • The current method for generating transgenic animals leads to unpredictable integration and expression.
  • Mosaicism in founder animals is a consequence of integration during DNA replication.
  • Strategies are needed to overcome challenges associated with gene expression variability in commercial applications.

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