An Improved Tet-On System for Gene Expression in Neurons Delivered by a Single Lentiviral Vector
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.
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Recent studies successfully applied V16 to control transgene expression in mice and non-human primates.
“Recent studies successfully applied V16 to control transgene expression in mice [85, 86] and non-human primates [62, 87].”
From Tet-On Systems For Doxycycline-inducible Gene Expression (2016) · cites this paper as [86]
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- Authors
- Tian, X., Wang, G., Xu, Y., Wang, P., Chen, S., Yang, H., Gao, F., Xu, A., Cao, F., Jin, X., Manyande, A., & Tian, Y.
- Affiliations
- Tongji Hospital (CN)
- Shandong Provincial Hospital (CN)
- Children's Hospital of Chongqing Medical University (CN)
- Psychology Department, Thames Valley University, Brentford, Middx TW8 9GA, United Kingdom.
- Corresponding author
- Tian, Yuke
- ORCID
- Venue
- Human Gene Therapy
- Publisher
- SAGE Publications
- Published
- 2009-02
- Volume
- 20
- Issue
- 2
- Pages
- 113-123
- ISSN
- 1043-0342 (ISSN-L)
- 1557-7422
- OpenAlex
- W2054724065
- Cited by
- 21
- References
- 38
- Citations by year
- 2021: 1 · 2020: 1 · 2019: 3 · 2017: 2 · 2016: 3 · 2015: 2 · 2014: 2 · 2013: 2 · 2012: 1
- FWCI
- 0.97
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Human Gene Therapy (publishedVersion)
- PubMed (publishedVersion)
- UWL Repository (University of West London) (submittedVersion)
- Funders
- National Natural Science Foundation of China
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Keywords
- Tet-On system, tetracycline-regulated gene expression, lentiviral vectors, neuron-specific enolase promoter, neuronal gene expression, doxycycline regulation
- MeSH terms
- Animals, Cell Line, Chickens, Electrophoresis, Polyacrylamide Gel, Gene Expression Regulation, HeLa Cells, Humans, Immunohistochemistry, Luciferases, Neurons, Phosphopyruvate Hydratase, Promoter Regions, Genetic, Protein Synthesis Inhibitors, Tetracycline, Blotting, Western, Lentivirus, Muromegalovirus, Models, Animal, Mice, Rats
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