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 Google Scholar
Google Scholar
Right arrow Articles by Schnepp, B. C.
Right arrow Articles by Johnson, P. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Schnepp, B. C.
Right arrow Articles by Johnson, P. R.

 Previous Article  |  Next Article 

Journal of Virology, February 2009, p. 1456-1464, Vol. 83, No. 3
0022-538X/09/$08.00+0     doi:10.1128/JVI.01686-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Infectious Molecular Clones of Adeno-Associated Virus Isolated Directly from Human Tissues{triangledown}

Bruce C. Schnepp,1 Ryan L. Jensen,1 K. Reed Clark,1,2,3 and Philip R. Johnson1,2,3*

The Research Institute, Nationwide Children's Hospital, Columbus, Ohio,1 Department of Pediatrics,2 Department of Molecular Immunology, Virology, and Medical Genetics, College of Medicine, Ohio State University, Columbus, Ohio3

Received 7 August 2008/ Accepted 14 November 2008

Adeno-associated virus (AAV) replication and biology have been extensively studied using cell culture systems, but there is precious little known about AAV biology in natural hosts. As part of our ongoing interest in the in vivo biology of AAV, we previously described the existence of extrachromosomal proviral AAV genomes in human tissues. In the current work, we describe the molecular structure of infectious DNA clones derived directly from these tissues. Sequence-specific linear rolling-circle amplification was utilized to isolate clones of native circular AAV DNA. Several molecular clones containing unit-length viral genomes directed the production of infectious wild-type AAV upon DNA transfection in the presence of adenovirus help. DNA sequence analysis of the molecular clones revealed the ubiquitous presence of a double-D inverted terminal repeat (ITR) structure, which implied a mechanism by which the virus is able to maintain ITR sequence continuity and persist in the absence of host chromosome integration. These data suggest that the natural life cycle of AAV, unlike that of retroviruses, might not have genome integration as an obligatory component.

* Corresponding author. Present address: Room 1216B, Abramson Research Center, The Children's Hospital of Philadelphia, 3615 Civic Center Blvd, Philadelphia, PA 19104-4318. Phone: (267) 426-0351. Fax: (267) 426-0363. E-mail: johnsonphi{at}chop.edu

{triangledown} Published ahead of print on 19 November 2008.

Journal of Virology, February 2009, p. 1456-1464, Vol. 83, No. 3
0022-538X/09/$08.00+0     doi:10.1128/JVI.01686-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»