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Efficient Generation of Recombinant Adenoviral Vectors by Cre-lox Recombination In Vitro

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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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Acknowledgments

We thank Ms. Donna Gschwend for secretarial assistance, Ms. Nancy Barrett for preparation of figures, and Ms. Kay Cherian for her helpful advice and comments. This work was supported in part by a grant from the National Institutes of Health (GM34902).

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Authors and Affiliations

  1. Department of Internal Medicine and Biological Chemistry, Howard Hughes Medical Institute, University of Michigan Medical Center, 1150 West Medical Center Drive, 4520 MSRB I, Ann Arbor, Michigan, 48109-0650, USA

    Kazunori Aoki, Christopher Barker &  Gary J. Nabel

  2. Department of Microbiology and Immunology and Comprehensive Cancer Center, University of Michigan Medical Center, Ann Arbor, Michigan, USA

    Michael J. Imperiale

  3. Research Service 1 51, VA Medical Center, 500 W. Fort Street, Boise, Idaho, USA

    Xavier Danthinne

Authors
  1. Kazunori Aoki
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  2. Christopher Barker
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  3. Xavier Danthinne
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  4. Michael J. Imperiale
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  5. Gary J. Nabel
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Corresponding author

Correspondence to Gary J. Nabel.

Additional information

Communicated by E. Nabel.

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Aoki, K., Barker, C., Danthinne, X. et al. Efficient Generation of Recombinant Adenoviral Vectors by Cre-lox Recombination In Vitro. Mol Med 5, 224–231 (1999). https://doi.org/10.1007/BF03402119

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  • DOI: https://doi.org/10.1007/BF03402119

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