Efficient generation of recombinant adenoviral vectors by Cre-lox recombination in vitro. |
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| Authors: | K Aoki C Barker X Danthinne M J Imperiale and G J Nabel |
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| Affiliation: | Howard Hughes Medical Institute, University of Michigan Medical Center, Department of Internal Medicine and Biological Chemistry, Ann Arbor 48109-0650, USA. |
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| Abstract: | BACKGROUND: Although recombinant adenovirus vectors are attractive for use in gene expression studies and therapeutic applications, the construction of these vectors remains relatively time-consuming. We report here a strategy that simplifies the production of adenoviruses using the Cre-loxP system. MATERIALS AND METHODS: Full-length recombinant adenovirus DNA was generated in vitro by Cre-mediated recombination between loxP sites in a linearized shuttle plasmid containing a transgene and adenovirus genomic DNA. RESULTS: After transfection of Cre-treated DNA into 293 cells, replication-defective viral vectors were rapidly obtained without detectable wild-type virus. CONCLUSION: This system facilitates the development of recombinant adenoviral vectors for basic and clinical research. |
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