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Journal of Virology, December 2002, p. 12900-12907, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12900-12907.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Conjugate-Based Targeting of Recombinant Adeno-Associated Virus Type 2 Vectors by Using Avidin-Linked Ligands

Selvarangan Ponnazhagan,1* Gandham Mahendra,1 Sanjay Kumar,1 John A. Thompson,2 and Mark Castillas, Jr.1

Departments of Pathology,1 Surgery, The University of Alabama at Birmingham, Birmingham, Alabama 352942

Received 19 July 2002/ Accepted 3 September 2002

The development of targeted vectors, capable of tissue-specific transduction, remains one of the important aspects of vector modification for gene therapy applications. Recombinant adeno-associated virus type 2 (rAAV-2)-based vectors are nonpathogenic, have relatively low immunogenicity, and are capable of long-term transgene expression. AAV-2 vectors bind primarily to heparan sulfate proteoglycan (HSPG), a receptor that is present in many tissues and cell types. Because of the widespread expression of HSPG on many tissues, targeted transduction in vivo appears to be limited with AAV-2 vectors. Thus, development of strategies to achieve transductional targeting will have a profound benefit in the future application of these vectors. We report here a novel conjugate-based targeting method to enhance tissue-specific transduction of AAV-2-based vectors. The present report utilized a high-affinity biotin-avidin interaction as a molecular bridge to cross-link purified targeting ligands, produced genetically as fusion proteins to core-streptavidin, in a prokaryotic expression system. Conjugation of the bispecific targeting protein to the vector was achieved by biotinylating purified rAAV-2 without abolishing the capsid structure, internalization, and subsequent transgene expression. The tropism-modified vectors, targeted via epidermal growth factor receptor (EGFR) or fibroblast growth factor 1{alpha} receptor (FGFR1{alpha}), resulted in a significant increase in transduction efficiency of EGFR-positive SKOV3.ip1 cells and FGFR1{alpha}-positive M07e cells, respectively. Further optimization of this method of targeting should enhance the potential of AAV-2 vectors in ex vivo and in vivo gene therapy and may form the basis for developing targeting methods for other AAV serotype capsids.

* Corresponding author. Mailing address: Department of Pathology, LHRB 513, The University of Alabama at Birmingham, 701 19th St. South, Birmingham, AL 35294-0007. Phone: (205) 934-6731. Fax: (205) 975-9927. E-mail: sponnazh{at}path.uab.edu.

Journal of Virology, December 2002, p. 12900-12907, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.12900-12907.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Daya, S., Berns, K. I. (2008). Gene Therapy Using Adeno-Associated Virus Vectors. Clin. Microbiol. Rev. 21: 583-593 [Abstract] [Full Text]  
  • Warrington, K. H. Jr., Gorbatyuk, O. S., Harrison, J. K., Opie, S. R., Zolotukhin, S., Muzyczka, N. (2004). Adeno-Associated Virus Type 2 VP2 Capsid Protein Is Nonessential and Can Tolerate Large Peptide Insertions at Its N Terminus. J. Virol. 78: 6595-6609 [Abstract] [Full Text]  
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