Abstract
Adeno-associated virus (AAV) gene delivery vectors are being investigated as vehicles for gene therapy for a wide variety of hereditary and acquired human diseases. AAV’s inability to self-propagate, ability to be maintained as an episome in the transduced cell, and relatively innocuous effects on the immune system make it the vector of choice for prolonged in vivo gene expression. AAV type 2 is the most commonly used serotype for gene delivery. AAV2 vectors will deliver DNA to a wide variety of cell types. The development of vectors derived from the other five serotypes has expanded the tissue tropism of the AAV vector system (1–6). Tropism depends on the presence of cell-surface receptor elements on the target cell. For AAV2, heparin sulfate proteoglycan (7), α2βV integrin (8) and the fibroblast growth factor receptor-1 (FGFR-1) (9) are believed to mediate the initial internalization steps in infection. The ubiquity of these cell-surface components confers a wide tropism on AAV2 vectors.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chiorini, J. A., Yang, L., Liu, Y., Safer, B., and Kotin, R. M. (1997) Cloning of adeno-associated virus type 4 (AAV4) and generation of recombinant AAV4 particles. J. Virol. 71, 6823–6833.
Chiorini, J. A., Kim, F., Yang, L., and Kotin, R. M. (1999) Cloning and characterization of adeno-associated virus type 5. J. Virol. 73, 1309–1319.
Duan, D., Yan, Z., Yue, Y, Ding, W., and Engelhardt, J. F. (2001) Enhancement of muscle gene delivery with pseudotyped adeno-associated virus type 5 correlates with myoblast differentiation. J. Virol. 75, 7662–7671.
Hildinger, M., Auricchio, A., Gao, G., Wang, L., Chirmule, N., and Wilson, J. M. (2001) Hybrid vectors based on adeno-associated virus serotypes 2 and 5 for muscle-directed gene transfer. J. Virol. 75, 6199–6203.
Rutledge, E. A., Halbert, C. L., and Russell, D. W. (1998) Infectious clones and vectors derived from adeno-associated virus (AAV) serotypes other than AAV type 2. J. Virol. 72, 309–319.
Xiao, W., Chirmule, N., Berta, S. C., McCullough, B., Gao, G., and Wilson, J. M. (1999) Gene therapy vectors based on adeno-associated virus type 1. J. Virol. 73, 3994–4003.
Summerford, C. and Samulski, R. J. (1998) Membrane-associated heparin sulfate proteoglycan is a receptor for adeno-associated virus type 2 virions. J. Virol 72, 1438–1445.
Summerford, C., Bartlett, J. S., and Samulski, R. J. (1999) AlphaV Beta5 integrin: a co-receptor for adeno-associated virus type 2 infection. Nat. Med. 5, 78–82.
Qing, K., Mah, C., Hansen, J., Zhou, S., Dwarki, V., and Srivastava, A. (1999) Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2. Nat. Med. 5, 71–77.
Ponnazhagan, S., Mukherjee, P., Wang, X. S., Qing, K., Kube, D. M., Mah, C., et al. (1997) Adeno-associated virus type 2-mediated transduction in primary human bone marrow-derived CD34+ hematopoietic progenitor cells: donor variation and correlation of transgene expression with cellular differentiation. J. Virol. 71, 8262–8267.
Duan, D., Yue, Y., Yan, Z., McCray, P. B., Jr., and Engelhardt, J. F. (1998) Polarity influences the efficiency of recombinant adenoassociated virus infection in differentiated airway epithelia. Hum. Gene Ther. 9, 2761–2776.
Zabner, J., Seiler, M., Walters, R., Kotin, R. M., Fulgeras, W., Davidson, B. L., and Chiorini, J. A. (2000) Adeno-associated virus type 5 (AAV5) but not AAV2 binds to the apical surfaces of airway epithelia and facilitates gene transfer. J. Virol. 74, 3852–3858.
Fisher, K. J., Gao, G.-P., Weitzman, M. D., DeMatteo, R., Burda, J. F., and Wilson, J. M. (1996) Transduction with recombinant adeno-associated virus for gene therapy is limited by leading-strand synthesis. J. Virol. 70, 520–532.
Alexander, I. E., Russell, D. W., and Miller, A. D. (1994) DNA-damaging agents greatly increase the transduction of nondividing cells by adeno-associated virus vectors. J. Virol. 68, 8282–8287.
Russell, D. W., Alexander, I. E., and Miller, A. D. (1995) DNA synthesis and topoi-somerase inhibitors increase transduction by adeno-associated virus vectors. Proc. Natl. Acad. Sci. USA 92, 5719–5723.
Ferrari, F. K., Samulski, T., Shenk, T., and Samulski, R. J. (1996) Second-strand synthesis is a rate-limiting step for efficient transduction by recombinant adeno-associated virus vectors. J. Virol. 70, 3227–3234.
Qing, K. Y., Wang, X.-S., Kube, D. M., Ponnazhagan, S., Bajpai, A., and Srivastava, A. (1997) Role of tyrosine phosphorylation of a cellular protein in adeno-associated virus 2-mediated transgene expression. Proc. Natl. Acad. Sci. USA 94, 10879–10884.
Mah, C., Qing, K., Khuntirat, B., Ponnazhagan, P., Wang, X.-S., Kube, D. M., et al. (1998) Adeno-associated virus type 2-mediated gene transfer: Role of epidermal growth factor receptor protein tyrosine kinase in transgene expression. J. Virol. 72, 9835–9843.
Douar, A.-M., Poulard, K., Stockholm, D., and Danos, O. (2001) Intracellular trafficking of adeno-assoicated virus vectors: routing to the late endosomal compartment and proteosome degradation. J. Virol. 75, 1824–1833.
Duan, D., Yue, Y., Yan, Z., and Engelhardt, J. F. (2000) Endosomal processing limits gene transfer to polarized airway epithelia by adeno-associated virus. J. Clin. Invest. 105, 1573–1587.
Collaco, R. F., Cao, X., and Trempe, J. P. (1999) A helper virus-free packaging system for recombinant adeno-associated virus vectors. Gene 238, 397–405.
Zeitlin, P. L., Lu, L., Rhim, J., Cutting, G., Stetten, G., Keiffer, K. A., et al. (1991) A cystic fibrosis bronchial epithelial cell line: immortalization by adeno-12-SV40 infection. Am. J. Resp. Cell Mol. Biol. 4, 313–319.
Zolotukhin, S., Byrne, B. J., Mason, E., Zolotukhin, I., Potter, M., Chesnut, K., et al. (1999) Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Ther. 6, 973–985.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2004 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Smith, A., Collaco, R., Trempe, J.P. (2004). AAV Vector Delivery to Cells in Culture. In: Heiser, W.C. (eds) Gene Delivery to Mammalian Cells. Methods in Molecular Biology™, vol 246. Humana Press. https://doi.org/10.1385/1-59259-650-9:167
Download citation
DOI: https://doi.org/10.1385/1-59259-650-9:167
Publisher Name: Humana Press
Print ISBN: 978-1-58829-095-3
Online ISBN: 978-1-59259-650-8
eBook Packages: Springer Protocols