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Mol Biol Med. 1989 Aug;6(4):283-98.

Mechanism of chromosomal integration of microinjected DNA.

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  • 1Department of Genetics, University of Edinburgh, Scotland.

Abstract

Most transgenic animals have been produced by directly injecting DNA into one of the embryo pronuclei in the period immediately following fertilization. Transgenic animals are produced when DNA becomes integrated into the chromosomes, in most cases the transgenic genotype is transmitted to progeny through the germ line. The characteristics of the foreign DNA include tandem arrangement of multiple copies of the input DNA, predominantly in direct rather than inverted orientation, and illegitimate recombination with the chromosome at the site of integration. Foreign DNA integrated into chromosomes of cultured cells has identical characteristics. We argue that these and other similarities between the integration of foreign DNA into the chromosomes of microinjected embryos on the one hand and on the other of transfected and microinjected cells, strongly suggest that the processes have the same basis. By considering mainly the literature relating to cell transfection and microinjection, we conclude that tandemly arranged concatemers of input foreign DNA are built up by a process of homologous but non-conservative recombination. End-joining and illegitimate recombination events characterize the integration of the concatemers into the chromosome, and also contribute to the formation of the concatemers. We also suggest that these superficially different processes are based on the same opportunistic repair-ligation mechanism; the frequencies of the different types of event reflect both the frequencies with which different sorts of association occur between DNA molecules and their relative stability.

PMID:
2695741
[PubMed - indexed for MEDLINE]
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