Efficient Integration of Short Interspersed Element-flanked Foreign DNA via Homologous Recombination

We investigated whether mouse short interspersed elements (SINEs) could influence the recombination frequency of foreign DNA. Vectors harboring a reporter gene in combinations of SINEs B1 and/or B2 or a portion of long interspersed element-1 were prepared and tested in vitro by a colony assay using...

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Bibliographic Details
Published in:The Journal of biological chemistry 1999-12, Vol.274 (51), p.36585-36591
Main Authors: Kang, Y K, Park, J S, Lee, C S, Yeom, Y I, Chung, A S, Lee, K K
Format: Article
Language:English
Subjects:
Animals
Base Sequence
DNA - genetics
Genetic Vectors
Mice
Molecular Sequence Data
Recombination, Genetic
short interspersed nuclear elements
Transfection
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Summary:We investigated whether mouse short interspersed elements (SINEs) could influence the recombination frequency of foreign DNA. Vectors harboring a reporter gene in combinations of SINEs B1 and/or B2 or a portion of long interspersed element-1 were prepared and tested in vitro by a colony assay using HC11 murine mammary epithelial cells and in vivo by microinjection into fetilized mouse eggs. In transfected HC11 cells, the number of colonies surviving G418 selection increased by 3.5-fold compared with control when the reporter was flanked by fused B1-B2 sequences. Similar results were obtained from microinjection study; in fetuses 11.5 days post coitum, transgene positives in control and SINE-flanked vectors were 16 and 53%, respectively. Individual B1- and B2-harboring vectors showed equivalent activities with each other, as determined by the colony assay (2.8-fold versus 3.2-fold compared with control). We determined the contribution of homologous recombination to the SINE-mediated increase in integration frequency through a polymerase chain reaction-based strategy; in more than half of embryos transgenes underwent homologous recombinations involving B1 sequences. These results demonstrate that the SINE sequences can increase the integration rate of foreign DNA and that such an increase is most likely due to the enhancement of homologous recombination.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.274.51.36585