Depot formation of doxycycline impairs Tet-regulated gene expression in vivo
Abstract
The tetracycline (Tet) system is widely used for regulation of gene expression in vitro and in vivo. We constructed C57BL/6 transgenic mice (rtTA-CM2) with strong and ubiquitous reverse transactivator (rtTA2(S)-M2) gene expression. rtTA-CM2 mice were crossed to Tet-responsive reporter mice (LC-1) conditionally expressing the firefly luciferase (FLuc) gene under control of a Tet-responsive element, which allowed sensitive quantification of the transactivator activity by bioluminescent imaging. Following doxycycline (dox) application, up to 10(5)-fold increase in BL signal was measured. rtTA activity was inducible in most analyzed organs. After dox withdrawal the BL signal decreased significantly but did not disappear completely, most likely due to a dox depot formation in vivo. The residual dox was sufficient to partly down-regulate a Tet-off controlled oncogene in a tumor transplantation experiment, resulting in reduced tumor growth. rtTA-CM2 mice may be a useful tool to analyze the function of genes in various organs but also reveal that down-regulation of gene expression is not complete.
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A more dox-sensitive rtTA variant, like V16, may demonstrate prolonged activity when the dox level drops after dox-withdrawal.
“Another obvious issue to take into consideration is that a more dox-sensitive rtTA variant, like V16, may demonstrate prolonged activity when the dox level drops after dox-withdrawal [84].”
From Tet-On Systems For Doxycycline-inducible Gene Expression (2016) · cites this paper as [84]
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References 18 · Cited by 13
- Authors
- Anders, K., Buschow, C., Charo, J., & Blankenstein, T.
- Affiliations
- Max Delbrück Center (DE)
- Charité – Universitätsmedizin Berlin (DE)
- Corresponding author
- Blankenstein, Thomas
- Venue
- Transgenic Research
- Published
- 2011-12-14
- Volume
- 21
- Issue
- 5
- Pages
- 1099-1107
- ISSN
- 0962-8819 (ISSN-L)
- 1573-9368
- OpenAlex
- W1970289038
- Cited by
- 13
- References
- 18
- Citations by year
- 2026: 1 · 2023: 1 · 2022: 1 · 2021: 1 · 2018: 1 · 2017: 1 · 2016: 3 · 2015: 1 · 2014: 1 · 2013: 1 · 2012: 1
- FWCI
- 0.60
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Transgenic Research (publishedVersion)
- PubMed (publishedVersion)
- MDC Repository (Max-Delbrueck-Center for Molecular Medicine) (submittedVersion)
- Funders
- Deutsche Forschungsgemeinschaft
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene Regulatory Network Analysis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Keywords
- tetracycline-inducible expression, bioluminescence imaging, tumor growth inhibition, conditional gene expression
- MeSH terms
- Animals, Antigens, Polyomavirus Transforming, Antineoplastic Agents, Crosses, Genetic, Doxycycline, Drug Screening Assays, Antitumor, Luciferases, Luminescent Measurements, Mice, Inbred C57BL, Mice, Transgenic, Microinjections, Oocytes, Promoter Regions, Genetic, Simian virus 40, Transcription, Genetic, Oncogene Proteins, Fusion, Trans-Activators, Gene Expression Regulation, Neoplastic, Founder Effect, Transgenes, Mice, Tandem Mass Spectrometry
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