Key result
Transfection of linear enhancerless SV40 DNA into monkey cells with integrated SV40 early regions resulted in up to 25% of cells producing wild-type virus via homologous recombination within 48 hours.
The SV40/COS cell system demonstrates a high frequency of homologous recombination between introduced and chromosomal DNA, providing a rapid assay for such recombination events.
Efficient homologous recombination occurs in mammalian cells in vitro; leaves open frequency, mechanisms, and therapeutic relevance in vivo.
We have detected a high frequency of homologous recombination between introduced and chromosomal DNA in mammalian cells. Linear enhancerless SV40 DNA has been transfected into monkey cells that have either one (COS1 cells) or five to seven (COS7 cells) copies of the SV40 early region stably integrated into their genome. Enhancer-containing wild-type SV40 DNA is formed as a result of homologous recombination of the introduced DNA with chromosomal DNA. Up to 25% of the successfully transfected cells produce wild-type virus within 48 hr after transfection. The highest levels of wild-type virus were produced from transfections of molecules that contained a double-strand break at positions of uninterrupted homology with the chromosomal template. This SV40/COS cell system provides a rapid assay for recombination between introduced and genomic DNA.
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Jasin et al. (1985) studied this question. Transfection of linear enhancerless SV40 DNA was evaluated on Production of wild-type virus via homologous recombination. Transfection of linear enhancerless SV40 DNA into monkey cells with integrated SV40 early regions resulted in up to 25% of cells producing wild-type virus via homologous recombination within 48 hours.
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