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Experimental Physiology 90.1 pp 33-37
DOI: 10.1113/expphysiol.2004.028209
© The Physiological Society 2005
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Symposium Report

Viral gene transfer in neuroscience: new tricks of the trade

Optimizing regulatable gene expression using adenoviral vectors

Youn-Bok Lee1, Colin P. J. Glover1, A. Siobhan Cosgrave1, Alison Bienemann1 and James B. Uney1

1 Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology (LINE), Bristol University, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

Abstract

Inducible gene expression systems have typically encountered limitations, such as pleitropic effects of the inducer, basal leakiness, toxicity of inducing agents and low levels of expression. However, recently non-toxic, tightly regulated control of transgene expression has been reported for several systems, the most frequently cited being the tetracycline gene control system. We have found that the individual components of the Tet system [the Tet transactivators and tetracycline responsive element (TRE)] function optimally to control gene expression when they are incorporated into separate adenoviral vectors. Furthermore, incorporation of the Woodchuck hepatitis virus post-transcriptional enhancer (WPRE) allows a dual vector Tet-regulatable Ad system to be used at very low titres (2 x 104) that elicit a minimal inflammatory response, with no loss of transgene expression or ability to regulate transgene expression. This and similar regulatable systems will benefit studies investigating neuronal gene function and those seeking to develop effective neuronal gene therapy strategies.

(Received 6 October 2004; accepted after revision 3 November 2004; first published online 12 November 2004)
Corresponding author J. B. Uney: Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology (LINE), Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK. Email: James.uney{at}bris.ac.uk






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