An improved Tet-On System for gene expression in neurons delivered by a single lentiviral vector

Tian, Xuebi, Wang, Gongming, Xu, Ying, Wang, Ping, Chen, Shasha, Yang, Hui, Gao, Feng, Xu, Aijun, Cao, Fei, Jin, Xiaogao, Manyande, Anne and Tian, Yu-Ke (2009) An improved Tet-On System for gene expression in neurons delivered by a single lentiviral vector. Human Gene Therapy, 20 (2). pp. 113-123. ISSN 1043-0342

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Abstract

In most cases, the successful application of gene therapy requires the development of vectors that can provide regulated control of therapeutic gene expression. We have reconstituted the Tet-On (tetracycline-regulated transgene expression) two-component system in a single lentiviral vector with insertion of a chicken chromatin insulator (cHS4) element between the two expression cassettes. Optimization of this vector included an improved reverse tetracycline-dependent trans-activator (rtTA) sequence developed through HIV viral evolution, and an rtTA-dependent, Tet-responsive element containing modifications of the tetO sequence (TRE-tight1) to improve leaky basal transcription. Transduction of HeLa cells with these lentiviral vectors resulted in a high level of rtTA expression in the presence of doxycycline. In neuronal cells, rtTA expression driven by a neuron-specific enolase (NSE) promoter was more efficient than gene expression from a murine cytomegalovirus promoter. Transgene expression from the NSE promoter also provided tightly regulated gene expression in neurons in vivo.

Item Type: Article
Subjects: Medicine and health > Microbiology
Depositing User: Anne Manyande
Date Deposited: 10 Jun 2014 13:21
Last Modified: 09 May 2017 10:58
URI: http://repository.uwl.ac.uk/id/eprint/889

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