JVI Figure table search 04
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Steinwaerder, D. S.
Right arrow Articles by Lieber, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Steinwaerder, D. S.
Right arrow Articles by Lieber, A.

 Previous Article  |  Next Article 

Journal of Virology, November 1999, p. 9303-9313, Vol. 73, No. 11
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Generation of Adenovirus Vectors Devoid of All Viral Genes by Recombination between Inverted Repeats

Dirk S. Steinwaerder, Cheryl A. Carlson, and André Lieber*

Division of Medical Genetics, University of Washington, Seattle, Washington 98195

Received 3 June 1999/Accepted 15 August 1999

Direct or inverse repeated sequences are important functional features of prokaryotic and eukaryotic genomes. Considering the unique mechanism, involving single-stranded genomic intermediates, by which adenovirus (Ad) replicates its genome, we investigated whether repetitive homologous sequences inserted into E1-deleted adenoviral vectors would affect replication of viral DNA. In these studies we found that inverted repeats (IRs) inserted into the E1 region could mediate predictable genomic rearrangements, resulting in vector genomes devoid of all viral genes. These genomes (termed Delta Ad.IR) contained only the transgene cassette flanked on both sides by precisely duplicated IRs, Ad packaging signals, and Ad inverted terminal repeat sequences. Generation of Delta Ad.IR genomes could also be achieved by coinfecting two viruses, each providing one inverse homology element. The formation of Delta Ad.IR genomes required Ad DNA replication and appeared to involve recombination between the homologous inverted sequences. The formation of Delta Ad.IR genomes did not depend on the sequence within or adjacent to the inverted repeat elements. The small Delta Ad.IR vector genomes were efficiently packaged into functional Ad particles. All functions for Delta Ad.IR replication and packaging were provided by the full-length genome amplified in the same cell. Delta Ad.IR vectors were produced at a yield of ~104 particles per cell, which could be separated from virions with full-length genomes based on their lighter buoyant density. Delta Ad.IR vectors infected cultured cells with the same efficiency as first-generation vectors; however, transgene expression was only transient due to the instability of deleted genomes within transduced cells. The finding that IRs present within Ad vector genomes can mediate precise genetic rearrangements has important implications for the development of new vectors for gene therapy approaches.

* Corresponding author. Mailing address: Division of Medical Genetics, Box 357720, University of Washington, Seattle, WA 98195. Phone: (206) 221-3973. Fax: (206) 685-8675. E-mail: lieber00{at}u.washington.edu.

Journal of Virology, November 1999, p. 9303-9313, Vol. 73, No. 11
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
J. Bacteriol. Mol. Cell. Biol. Microbiol. Mol. Biol. Rev.
Clin. Vaccine Immunol. ALL ASM JOURNALS

Copyright © 1999 by the American Society for Microbiology. All rights reserved.