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 Logan, A. C.
Right arrow Articles by Kohn, D. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Logan, A. C.
Right arrow Articles by Kohn, D. B.

 Previous Article  |  Next Article 

Journal of Virology, August 2004, p. 8421-8436, Vol. 78, No. 16
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.16.8421-8436.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Integrated Self-Inactivating Lentiviral Vectors Produce Full-Length Genomic Transcripts Competent for Encapsidation and Integration

Aaron C. Logan,1,2 Dennis L. Haas,1,2 Tal Kafri,3 and Donald B. Kohn1,2*

Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California,1 Division of Research Immunology and Bone Marrow Transplantation, Children's Hospital Los Angeles, Los Angeles, California,2 Department of Microbiology and Immunology, University of North Carolina Chapel Hill, Chapel Hill, North Carolina3

Received 6 February 2004/ Accepted 9 April 2004

To make human immunodeficiency virus type 1 (HIV-1)-based vectors safer for use in the research and clinical setting, a significant modification to the HIV-1 genome has been the deletion of promoter and enhancer elements from the U3 region of the long terminal repeat (LTR). Vectors containing this deletion are thought to have no LTR-directed transcription and are called self-inactivating (SIN) lentivectors. Using four distinct approaches, we show that SIN lentivectors continue to have promoter activity near the 5' LTR, which is responsible for the production of full-length vector transcripts. To verify that transcripts derived from the LTR in SIN lentivectors are competent for encapsidation and integration, we transduced a lentiviral packaging cell line with a SIN lentivector and then observed the production of viable vector particles containing full-length SIN lentivector genomes. We have also attempted to identify sequences in the SIN lentivector which are responsible for transcriptional activation at the 5' LTR. Using different segments of the vector LTR and leader region in a promoter assay, we have determined that the residual promoter activity is contained entirely within the leader region and that, although this element is downstream of the transcription initiation site, it is capable of initiating transcription from the 5' end of R in the LTR. Mutation of leader region binding sites for the transcriptional activators downstream binding factor 1 (DBF1) and SP1 reduces transcription from the SIN LTR by up to 80%. Knowledge of the potential for mobilization of HIV-1-derived SIN lentivectors will be important for the design of future gene therapy trials with such vectors.

* Corresponding author. Mailing address: Children's Hospital Los Angeles, Division of Research Immunology/Bone Marrow Transplantation, 4650 Sunset Blvd., MS62, Los Angeles, CA 90027-6016. Phone: (323) 669-4617. Fax: (323) 667-1021. E-mail: dkohn{at}chla.usc.edu.

Journal of Virology, August 2004, p. 8421-8436, Vol. 78, No. 16
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.16.8421-8436.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hendrickson, B., Senadheera, D., Mishra, S., Bui, K. C. T., Wang, X., Chan, B., Petersen, D., Pepper, K., Lutzko, C. (2007). Development of Lentiviral Vectors with Regulated Respiratory Epithelial Expression In Vivo. Am. J. Respir. Cell Mol. Bio. 37: 414-423 [Abstract] [Full Text]  
  • Poon, B., Chang, M. A., Chen, I. S. Y. (2007). Vpr Is Required for Efficient Nef Expression from Unintegrated Human Immunodeficiency Virus Type 1 DNA. J. Virol. 81: 10515-10523 [Abstract] [Full Text]  
  • Kraunus, J., Zychlinski, D., Heise, T., Galla, M., Bohne, J., Baum, C. (2006). Murine Leukemia Virus Regulates Alternative Splicing through Sequences Upstream of the 5{middle dot} Splice Site. J. Biol. Chem. 281: 37381-37390 [Abstract] [Full Text]  
  • Lucke, S., Grunwald, T., Uberla, K. (2005). Reduced Mobilization of Rev-Responsive Element-Deficient Lentiviral Vectors. J. Virol. 79: 9359-9362 [Abstract] [Full Text]  
  • Hanawa, H., Persons, D. A., Nienhuis, A. W. (2005). Mobilization and Mechanism of Transcription of Integrated Self-Inactivating Lentiviral Vectors. J. Virol. 79: 8410-8421 [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»