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Capture of a recombination activating sequence from mammalian cells

Gene Therapy volume 6, pages 1819–1825 (1999)Cite this article

Abstract

We have developed a genetic trap for identifying sequences that promote homologous DNA recombination. The trap employs a retroviral vector that normally disables itself after one round of replication. Insertion of defined DNA sequences into the vector induced the repair of a 300 base pair deletion, which restored its ability to replicate. Tests of random sequence libraries made in the vector revealed a putative recombination signal (CCCACCC). When this heptamer or an abbreviated form (CCCACC) were reinserted into the vector, they stimulated vector repair and other DNA rearrangements. Mutant forms of these oligomers (eg CCCAACC or CCWACWS) did not. Our data suggest that the recombination events occurred within 48 h after transfection.

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Acknowledgements

We would like to thank Hugh Fisher for his help in preparing this manuscript. We would like to thank Dr Howard Passmore (Rutgers University) for his critical and helpful comments on this paper. We also would like to thank Dr Ren Sun and Dr G Miller (Yale University) for communication before publication. This work was sponsored by grants from the March of Dimes Birth Defect Foundation and the New Jersey Commission on Cancer Research.

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  1. Division of Infectious Diseases, The Dorrance H Hamilton Laboratories, Center for Human Virology, Jefferson Medical College, Thomas Jefferson University, Jefferson Alumni Hall, 1020 Locust Street, Rm 329, Philadelphia, 19107, PA, USA

    P Olson & R Dornburg

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  2. R Dornburg
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Olson, P., Dornburg, R. Capture of a recombination activating sequence from mammalian cells. Gene Ther 6, 1819–1825 (1999). https://doi.org/10.1038/sj.gt.3301035

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