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 Vincent-Lacaze, N.
Right arrow Articles by Danos, O.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Vincent-Lacaze, N.
Right arrow Articles by Danos, O.

 Previous Article  |  Next Article 

Journal of Virology, March 1999, p. 1949-1955, Vol. 73, No. 3
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Structure of Adeno-Associated Virus Vector DNA following Transduction of the Skeletal Muscle

Nathalie Vincent-Lacaze,1 Richard O. Snyder,2 Régis Gluzman,1 Delphine Bohl,3 Catherine Lagarde,2 and Olivier Danos1,*

Gene Therapy Program, Genethon III, CNRS URA 1922, Evry,1 and Laboratoire Retrovirus et Transfert Génétique, Institut Pasteur, Paris,3 France, and Cell Genesys, Foster City, California2

Received 16 November 1998/Accepted 30 November 1998

The skeletal muscle provides a very permissive physiological environment for adeno-associated virus (AAV) type 2-mediated gene transfer. We have studied the early steps leading to the establishment of permanent transgene expression, after injection of recombinant AAV (rAAV) particles in the quadriceps muscle of mice. The animals received an rAAV encoding a secreted protein, murine erythropoietin (mEpo), under the control of the human cytomegalovirus major immediate-early promoter and were sacrificed between 1 and 60 days after injection. The measurement of plasma Epo levels and of hematocrits indicated a progressive increase of transgene expression over the first 2 weeks, followed by a stabilization at maximal plateau values. The rAAV sequences were analyzed by Southern blotting following neutral or alkaline gel electrophoresis of total DNA from injected muscles. While a high number of rAAV sequences were detected during the first 5 days following the injection, only a few percent of these sequences was retained in the animals analyzed after 2 weeks, in which transgene expression was maximal. Double-stranded DNA molecules resulting from de novo second-strand synthesis were detected as early as day 1, indicating that this crucial step of AAV-mediated gene transfer is readily accomplished in the muscle. The templates driving stable gene expression at later time points are low in copy number and structured as high-molecular-weight concatemers or interlocked circles. The presence of the circular form of the rAAV genomes at early time points suggests that the molecular transformations involved in the formation of stable concatemers may involve a rolling-circle type of DNA replication.

* Corresponding author. Mailing address: Gene Therapy Program, Genethon III, CNRS URA 1922, Evry, France. Phone: 33-1-69 47 29 64. Fax: 33-1-69 47 28 38. E-mail: odanos{at}genethon.fr.

Journal of Virology, March 1999, p. 1949-1955, Vol. 73, No. 3
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Tafuro, S., Ayuso, E., Zacchigna, S., Zentilin, L., Moimas, S., Dore, F., Giacca, M. (2009). Inducible adeno-associated virus vectors promote functional angiogenesis in adult organisms via regulated vascular endothelial growth factor expression. Cardiovasc Res 83: 663-671 [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]  
  • Goyenvalle, A., Vulin, A., Fougerousse, F., Leturcq, F., Kaplan, J.-C., Garcia, L., Danos, O. (2004). Rescue of Dystrophic Muscle Through U7 snRNA-Mediated Exon Skipping. Science 306: 1796-1799 [Abstract] [Full Text]  
  • Duan, D., Yue, Y., Engelhardt, J. F. (2003). Consequences of DNA-Dependent Protein Kinase Catalytic Subunit Deficiency on Recombinant Adeno-Associated Virus Genome Circularization and Heterodimerization in Muscle Tissue. J. Virol. 77: 4751-4759 [Abstract] [Full Text]  
  • Schnepp, B. C., Clark, K. R., Klemanski, D. L., Pacak, C. A., Johnson, P. R. (2003). Genetic Fate of Recombinant Adeno-Associated Virus Vector Genomes in Muscle. J. Virol. 77: 3495-3504 [Abstract] [Full Text]  
  • Wang, F., Rendahl, K. G., Manning, W. C., Quiroz, D., Coyne, M., Miller, S. S. (2003). AAV-Mediated Expression of Vascular Endothelial Growth Factor Induces Choroidal Neovascularization in Rat. IOVS 44: 781-790 [Abstract] [Full Text]  
  • Favre, D., Blouin, V., Provost, N., Spisek, R., Porrot, F., Bohl, D., Marme, F., Cherel, Y., Salvetti, A., Hurtrel, B., Heard, J.-M., Riviere, Y., Moullier, P. (2002). Lack of an Immune Response against the Tetracycline-Dependent Transactivator Correlates with Long-Term Doxycycline-Regulated Transgene Expression in Nonhuman Primates after Intramuscular Injection of Recombinant Adeno-Associated Virus. J. Virol. 76: 11605-11611 [Abstract] [Full Text]  
  • Chamberlain, J. S. (2002). Gene therapy of muscular dystrophy. Hum Mol Genet 11: 2355-2362 [Abstract] [Full Text]  
  • Aikawa, R., Huggins, G. S., Snyder, R. O. (2002). Cardiomyocyte-specific Gene Expression Following Recombinant Adeno-associated Viral Vector Transduction. J. Biol. Chem. 277: 18979-18985 [Abstract] [Full Text]  
  • Bantel-Schaal, U., Hub, B., Kartenbeck, J. (2002). Endocytosis of Adeno-Associated Virus Type 5 Leads to Accumulation of Virus Particles in the Golgi Compartment. J. Virol. 76: 2340-2349 [Abstract] [Full Text]  
  • Zentilin, L., Marcello, A., Giacca, M. (2001). Involvement of Cellular Double-Stranded DNA Break Binding Proteins in Processing of the Recombinant Adeno-Associated Virus Genome. J. Virol. 75: 12279-12287 [Abstract] [Full Text]  
  • Raben, N., Lu, N., Nagaraju, K., Rivera, Y., Lee, A., Yan, B., Byrne, B., Meikle, P.J., Umapathysivam, K., Hopwood, J.J., Plotz, P.H. (2001). Conditional tissue-specific expression of the acid {alpha}-glucosidase (GAA) gene in the GAA knockout mice: implications for therapy. Hum Mol Genet 10: 2039-2047 [Abstract] [Full Text]  
  • Ponnazhagan, S., Curiel, D. T., Shaw, D. R., Alvarez, R. D., Siegal, G. P. (2001). Adeno-associated Virus for Cancer Gene Therapy. Cancer Res. 61: 6313-6321 [Full Text]  
  • Nakai, H., Yant, S. R., Storm, T. A., Fuess, S., Meuse, L., Kay, M. A. (2001). Extrachromosomal Recombinant Adeno-Associated Virus Vector Genomes Are Primarily Responsible for Stable Liver Transduction In Vivo. J. Virol. 75: 6969-6976 [Abstract] [Full Text]  
  • Sarukhan, A., Camugli, S., Gjata, B., von Boehmer, H., Danos, O., Jooss, K. (2001). Successful Interference with Cellular Immune Responses to Immunogenic Proteins Encoded by Recombinant Viral Vectors. J. Virol. 75: 269-277 [Abstract] [Full Text]  
  • Su, H., Lu, R., Kan, Y. W. (2000). Adeno-associated viral vector-mediated vascular endothelial growth factor gene transfer induces neovascular formation in ischemic heart. Proc. Natl. Acad. Sci. USA 10.1073/pnas.250488097v1 [Abstract] [Full Text]  
  • Nakai, H., Storm, T. A., Kay, M. A. (2000). Recruitment of Single-Stranded Recombinant Adeno-Associated Virus Vector Genomes and Intermolecular Recombination Are Responsible for Stable Transduction of Liver In Vivo. J. Virol. 74: 9451-9463 [Abstract] [Full Text]  
  • Haberman, R. P., McCown, T. J., Samulski, R. J. (2000). Novel Transcriptional Regulatory Signals in the Adeno-Associated Virus Terminal Repeat A/D Junction Element. J. Virol. 74: 8732-8739 [Abstract] [Full Text]  
  • Tattersall, P. (2000). The persistence of alien genomes. Proc. Natl. Acad. Sci. USA 97: 6239-6241 [Full Text]  
  • Malik, A. K., Monahan, P. E., Allen, D. L., Chen, B.-G., Samulski, R. J., Kurachi, K. (2000). Kinetics of Recombinant Adeno-Associated Virus-Mediated Gene Transfer. J. Virol. 74: 3555-3565 [Abstract] [Full Text]  
  • Miao, C. H., Nakai, H., Thompson, A. R., Storm, T. A., Chiu, W., Snyder, R. O., Kay, M. A. (2000). Nonrandom Transduction of Recombinant Adeno-Associated Virus Vectors in Mouse Hepatocytes In Vivo: Cell Cycling Does Not Influence Hepatocyte Transduction. J. Virol. 74: 3793-3803 [Abstract] [Full Text]  
  • Yang, J., Zhou, W., Zhang, Y., Zidon, T., Ritchie, T., Engelhardt, J. F. (1999). Concatamerization of Adeno-Associated Virus Circular Genomes Occurs through Intermolecular Recombination. J. Virol. 73: 9468-9477 [Abstract] [Full Text]  
  • Su, H., Lu, R., Kan, Y. W. (2000). Adeno-associated viral vector-mediated vascular endothelial growth factor gene transfer induces neovascular formation in ischemic heart. Proc. Natl. Acad. Sci. USA 97: 13801-13806 [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»