Skip Navigation

Nucleic Acids Research 2005 33(6):e63; doi:10.1093/nar/gni062
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (571K) Freely available
Right arrow Screen PDF (244K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
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 Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Markusic, D.
Right arrow Articles by Seppen, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Markusic, D.
Right arrow Articles by Seppen, J.
Related Collections
Right arrow Recombinant DNA expression
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 4 April 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Methods Online

Comparison of single regulated lentiviral vectors with rtTA expression driven by an autoregulatory loop or a constitutive promoter

David Markusic*, Ronald Oude-Elferink, Atze T. Das1, Ben Berkhout1 and Jurgen Seppen

AMC Liver Center S1-172, Meibergdreef 69, 1105BK Amsterdam, The Netherlands 1Department of Human Retrovirology, Academic Medical Center, University of Amsterdam Meibergdreef 15, 1105AZ Amsterdam, The Netherlands

*To whom correspondence should be addressed. Tel: +31 205668701; Fax: +31 205669190; Email: d.m.markusic{at}amc.uva.nl

Received December 23, 2004. Revised March 14, 2005. Accepted March 14, 2005.

Regulated expression of a therapeutic gene is crucial for safe and efficacious gene therapy. Many inducible regulatory systems use a constitutive promoter to express a regulatory protein, such as rtTA in the Tet-On system, which may restrict their use because of cytotoxicity and immunogenicity. Autoregulatory expression of rtTA provides extremely low levels of rtTA when transgene expression is off, with rapid transgene induction upon addition of doxycycline. Lentiviral vectors efficiently transfer genes to dividing and non-dividing cells with long-term gene expression both in vitro and in vivo. We compared regulatory function in a single lentiviral vector where rtTA was either expressed from a constitutive promoter or placed in an autoregulatory loop. Autoregulatory expression of rtTA was superior to constitutive promoter expression, resulting in higher viral titers, undetectable levels of both rtTA and transgene expression in the absence of doxycycline, improved induction kinetics and increased induction levels in all cells tested. We further expanded the utility of the autoregulatory vector by using an improved rtTA variant with an increased sensitivity to doxycycline. This lentiviral vector with doxycycline-regulated transgene expression may be useful for gene therapy applications and in experimental settings where strict temporal expression of a transgene is required.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Proc. Natl. Acad. Sci. USAHome page
K.-J. Shin, E. A. Wall, J. R. Zavzavadjian, L. A. Santat, J. Liu, J.-I. Hwang, R. Rebres, T. Roach, W. Seaman, M. I. Simon, et al.
A single lentiviral vector platform for microRNA-based conditional RNA interference and coordinated transgene expression
PNAS, September 12, 2006; 103(37): 13759 - 13764.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
W. A. Kues, R. Schwinzer, D. Wirth, E. Verhoeyen, E. Lemme, D. Herrmann, B. Barg-Kues, H. Hauser, K. Wonigeit, and H. Niemann
Epigenetic silencing and tissue independent expression of a novel tetracycline inducible system in double-transgenic pigs
FASEB J, June 1, 2006; 20(8): 1200 - 1202.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.