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 Nakai, H.
Right arrow Articles by Kay, M. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nakai, H.
Right arrow Articles by Kay, M. A.
Right arrowPubmed/NCBI databases
*Protein
Medline Plus Health Information
*Genes and Gene Therapy

 Previous Article  |  Next Article 

Journal of Virology, March 2005, p. 3606-3614, Vol. 79, No. 6
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.6.3606-3614.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Large-Scale Molecular Characterization of Adeno-Associated Virus Vector Integration in Mouse Liver

Hiroyuki Nakai,1* Xiaolin Wu,2 Sally Fuess,1 Theresa A. Storm,1,3 David Munroe,2 Eugenio Montini,4,{dagger} Shawn M. Burgess,5 Markus Grompe,4,6 and Mark A. Kay1,3

Departments of Pediatrics,1 Genetics, Stanford University School of Medicine, Stanford, California,3 Laboratory of Molecular Technology, SAIC-Frederick, National Cancer Institute-Frederick, Frederick,2 Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland,5 Departments of Molecular and Medical Genetics,4 Pediatrics, Oregon Health & Science University, Portland, Oregon6

Received 24 June 2004/ Accepted 29 October 2004

Recombinant adeno-associated virus (rAAV) vector holds promise for gene therapy. Despite a low frequency of chromosomal integration of vector genomes, recent studies have raised concerns about the risk of rAAV integration because integration occurs preferentially in genes and accompanies chromosomal deletions, which may lead to loss-of-function insertional mutagenesis. Here, by analyzing 347 rAAV integrations in mice, we elucidate novel features of rAAV integration: the presence of hot spots for integration and a strong preference for integrating near gene regulatory sequences. The most prominent hot spot was a harmless chromosomal niche in the rRNA gene repeats, whereas nearly half of the integrations landed near transcription start sites or CpG islands, suggesting the possibility of activating flanking cellular disease genes by vector integration, similar to retroviral gain-of-function insertional mutagenesis. Possible cancer-related genes were hit by rAAV integration at a frequency of 3.5%. In addition, the information about chromosomal changes at 218 integration sites and 602 breakpoints of vector genomes have provided a clue to how vector terminal repeats and host chromosomal DNA are joined in the integration process. Thus, the present study provides new insights into the risk of rAAV-mediated insertional mutagenesis and the mechanisms of rAAV integration.

* Corresponding author. Mailing address: Department of Pediatrics, 300 Pasteur Dr., Grant Bldg., Rm. S374, Stanford University School of Medicine, Stanford, CA 94305. Phone: (650) 725-7487. Fax: (650) 736-2068. E-mail: nakaih{at}stanford.edu.

{dagger} Present address: Istituto per la Ricerca e la Cura del Cancro, Candiolo, Turin, Italy.

Journal of Virology, March 2005, p. 3606-3614, Vol. 79, No. 6
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.6.3606-3614.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Niemeyer, G. P., Herzog, R. W., Mount, J., Arruda, V. R., Tillson, D. M., Hathcock, J., van Ginkel, F. W., High, K. A., Lothrop, C. D. Jr (2009). Long-term correction of inhibitor-prone hemophilia B dogs treated with liver-directed AAV2-mediated factor IX gene therapy. Blood 113: 797-806 [Abstract] [Full Text]  
  • Davis, J., Westfall, M. V., Townsend, D., Blankinship, M., Herron, T. J., Guerrero-Serna, G., Wang, W., Devaney, E., Metzger, J. M. (2008). Designing Heart Performance by Gene Transfer. Physiol. Rev. 88: 1567-1651 [Abstract] [Full Text]  
  • Inagaki, K., Piao, C., Kotchey, N. M., Wu, X., Nakai, H. (2008). Frequency and Spectrum of Genomic Integration of Recombinant Adeno-Associated Virus Serotype 8 Vector in Neonatal Mouse Liver. J. Virol. 82: 9513-9524 [Abstract] [Full Text]  
  • Penaud-Budloo, M., Le Guiner, C., Nowrouzi, A., Toromanoff, A., Cherel, Y., Chenuaud, P., Schmidt, M., von Kalle, C., Rolling, F., Moullier, P., Snyder, R. O. (2008). Adeno-Associated Virus Vector Genomes Persist as Episomal Chromatin in Primate Muscle. J. Virol. 82: 7875-7885 [Abstract] [Full Text]  
  • Inagaki, K., Ma, C., Storm, T. A., Kay, M. A., Nakai, H. (2007). The Role of DNA-PKcs and Artemis in Opening Viral DNA Hairpin Termini in Various Tissues in Mice. J. Virol. 81: 11304-11321 [Abstract] [Full Text]  
  • Inagaki, K., Lewis, S. M., Wu, X., Ma, C., Munroe, D. J., Fuess, S., Storm, T. A., Kay, M. A., Nakai, H. (2007). DNA Palindromes with a Modest Arm Length of {gtrsim}20 Base Pairs Are a Significant Target for Recombinant Adeno-Associated Virus Vector Integration in the Liver, Muscles, and Heart in Mice. J. Virol. 81: 11290-11303 [Abstract] [Full Text]  
  • Wang, J., Xie, J., Lu, H., Chen, L., Hauck, B., Samulski, R. J., Xiao, W. (2007). Existence of transient functional double-stranded DNA intermediates during recombinant AAV transduction. Proc. Natl. Acad. Sci. USA 104: 13104-13109 [Abstract] [Full Text]  
  • Donsante, A., Miller, D. G., Li, Y., Vogler, C., Brunt, E. M., Russell, D. W., Sands, M. S. (2007). AAV Vector Integration Sites in Mouse Hepatocellular Carcinoma. Science 317: 477-477 [Abstract] [Full Text]  
  • Drew, H. R., Lockett, L. J., Both, G. W. (2007). Increased complexity of wild-type adeno-associated virus-chromosomal junctions as determined by analysis of unselected cellular genomes. J. Gen. Virol. 88: 1722-1732 [Abstract] [Full Text]  
  • Dunbar, C. E. (2007). The Yin and Yang of Stem Cell Gene Therapy: Insights into Hematopoiesis, Leukemogenesis, and Gene Therapy Safety. ASH Education Book 2007: 460-465 [Abstract] [Full Text]  
  • Vasileva, A., Linden, R. M., Jessberger, R. (2006). Homologous recombination is required for AAV-mediated gene targeting. Nucleic Acids Res 34: 3345-3360 [Abstract] [Full Text]  
  • Kuck, D., Lau, T., Leuchs, B., Kern, A., Muller, M., Gissmann, L., Kleinschmidt, J. A. (2006). Intranasal Vaccination with Recombinant Adeno-Associated Virus Type 5 against Human Papillomavirus Type 16 L1. J. Virol. 80: 2621-2630 [Abstract] [Full Text]  
  • Grimm, D., Pandey, K., Nakai, H., Storm, T. A., Kay, M. A. (2006). Liver Transduction with Recombinant Adeno-Associated Virus Is Primarily Restricted by Capsid Serotype Not Vector Genotype. J. Virol. 80: 426-439 [Abstract] [Full Text]  
  • Ohbayashi, F., Balamotis, M. A., Kishimoto, A., Aizawa, E., Diaz, A., Hasty, P., Graham, F. L., Caskey, C. T., Mitani, K. (2005). Correction of chromosomal mutation and random integration in embryonic stem cells with helper-dependent adenoviral vectors. Proc. Natl. Acad. Sci. USA 102: 13628-13633 [Abstract] [Full Text]  


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