This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kubo, S. Articles by Mitani, K. Search for Related Content PubMed PubMed Citation Articles by Kubo, S. Articles by Mitani, K.

 Previous Article  |  Next Article 

Journal of Virology, March 2003, p. 2964-2971, Vol. 77, No. 5
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.5.2964-2971.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

A New Hybrid System Capable of Efficient Lentiviral Vector Production and Stable Gene Transfer Mediated by a Single Helper-Dependent Adenoviral Vector

Shuji Kubo¹ and Kohnosuke Mitani^1,2*

Department of Microbiology, Immunology and Molecular Genetics, UCLA School of Medicine,¹ Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, California 90095-1747²

Received 9 September 2002/ Accepted 3 December 2002

To achieve efficient and sustained gene expression, we developed a new lentivirus/adenovirus hybrid vector (LA vector) that encodes sequences required for production of a human immunodeficiency virus-based lentiviral vector (i.e., a lentiviral vector, a gag/pol/rev expression cassette, a tetracycline-inducible envelope cassette, and the tetracycline-inducible transcriptional activator cassette) in a single helper-dependent adenovirus vector backbone. Via either transfection or infection, human cell lines transduced with the LA vector produced a lentiviral vector in a doxycycline-dependent manner at titers up to 10⁵ to 10⁶ green fluorescent protein transducing units per ml, which are comparable to the titers obtained by conventional multiple plasmid transfection methods. Efficient spread and persistent expression of the transgene were observed in cells maintained in long-term culture that had been infected with the LA vector. Furthermore, when cocultured with adherent cells infected with the LA vector, the human T-cell leukemia cell line was successfully transduced with a marker gene. This LA vector possesses the advantages of efficient gene transfer from an adenoviral vector and stable integration from a lentiviral vector; therefore, it might have potential for a variety of gene therapy applications.

---

* Corresponding author. Mailing address: Department of Microbiology, Immunology and Molecular Genetics, UCLA School of Medicine, Box 951781, Los Angeles, CA 90095-1781. Phone: (310) 267-2031. Fax: (310) 206-5553. E-mail: mitani{at}ucla.edu.

---

Journal of Virology, March 2003, p. 2964-2971, Vol. 77, No. 5
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.5.2964-2971.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Kubo, S., Seleme, M. d. C., Soifer, H. S., Perez, J. L. G., Moran, J. V., Kazazian, H. H. Jr., Kasahara, N. (2006). L1 retrotransposition in nondividing and primary human somatic cells. Proc. Natl. Acad. Sci. USA 103: 8036-8041 [Abstract] [Full Text]  
• Wang, H., Shayakhmetov, D. M., Leege, T., Harkey, M., Li, Q., Papayannopoulou, T., Stamatoyannopolous, G., Lieber, A. (2005). A Capsid-Modified Helper-Dependent Adenovirus Vector Containing the {beta}-Globin Locus Control Region Displays a Nonrandom Integration Pattern and Allows Stable, Erythroid-Specific Gene Expression. J. Virol. 79: 10999-11013 [Abstract] [Full Text]  
• Mitta, B., Weber, C. C., Rimann, M., Fussenegger, M. (2004). Design and in vivo characterization of self-inactivating human and non-human lentiviral expression vectors engineered for streptogramin-adjustable transgene expression. Nucleic Acids Res 32: e106-e106 [Abstract] [Full Text]