46 results match your criteria tet-controlled


Recent evolution of a TET-controlled and DPPA3/STELLA-driven pathway of passive DNA demethylation in mammals.

Nat Commun 2020 11 24;11(1):5972. Epub 2020 Nov 24.

Department of Biology II and Center for Integrated Protein Science Munich (CIPSM), Human Biology and BioImaging, Ludwig-Maximilians-Universität München, Planegg-Martinsried, Germany.

Genome-wide DNA demethylation is a unique feature of mammalian development and naïve pluripotent stem cells. Here, we describe a recently evolved pathway in which global hypomethylation is achieved by the coupling of active and passive demethylation. TET activity is required, albeit indirectly, for global demethylation, which mostly occurs at sites devoid of TET binding. Read More

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November 2020

Generation and Characterization of Inducible Lung and Skin-Specific IL-22 Transgenic Mice.

Methods Mol Biol 2021 ;2223:115-132

Department of Molecular Microbiology and Immunology and Department of Pediatrics, Brown University Warren Alpert Medical School, Providence, RI, USA.

IL-22 is an IL-10 family cytokine that is increased in asthma and atopic dermatitis (AD). However, the specific role of IL-22 in the pathogenesis of allergic lung inflammation and AD in vivo has yet to be elucidated. We aimed to develop mouse models of allergic diseases in the lung and skin with inducible and tissue-specific expression of IL-22, using a tetracycline (Tet)-controlled system. Read More

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Optimisation of Tet-On inducible systems for Sleeping Beauty-based chimeric antigen receptor (CAR) applications.

Sci Rep 2020 08 4;10(1):13125. Epub 2020 Aug 4.

Department of Microbiology and Immunology, University of Otago, Dunedin, Otago, 9010, New Zealand.

Regulated expression of genetic elements that either encode polypeptides or various types of functional RNA is a fundamental goal for gene therapy. Inducible expression may be preferred over constitutive promoters to allow clinician-based control of gene expression. Existing Tet-On systems represent one of the tightest rheostats for control of gene expression in mammals. Read More

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A Tet/Q Hybrid System for Robust and Versatile Control of Transgene Expression in C. elegans.

iScience 2019 Jan 27;11:224-237. Epub 2018 Dec 27.

School of Life Sciences and Technology, ShanghaiTech University, Shanghai, China; Department Immunobiology, Yale University School of Medicine, New Haven, CT, USA. Electronic address:

Binary gene regulatory tools such as the Tetracycline (Tet)-controlled transcription system have revolutionized genetic research in multiple organisms, but their applications to the worm remain very limited. Here we report that the canonical Tet system is largely inactive in the worm but can be adapted for the worm by introducing multiple modifications, a crucial one being the use of the transcription activation domain from the fungal Q binary system. The resultant Tet/Q hybrid system proves more robust and flexible than either of its precursors, enabling elaborate modes of transgene manipulation previously hard to achieve in the worm, including inducible intersectional regulation and, in combination with the Q system, independent control of distinct transgenes within the same cells. Read More

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January 2019

Tet-Inducible Production of Infectious Zika Virus from the Full-Length cDNA Clones of African- and Asian-Lineage Strains.

Viruses 2018 12 9;10(12). Epub 2018 Dec 9.

Department of Microbiology, University of Alabama at Birmingham School of Medicine, Birmingham, AL 35294, USA.

Zika virus (ZIKV) is a mosquito-borne flavivirus that has emerged as an important human viral pathogen, causing congenital malformation including microcephaly among infants born to mothers infected with the virus during pregnancy. Phylogenetic analysis suggested that ZIKV can be classified into African and Asian lineages. In this study, we have developed a stable plasmid-based reverse genetic system for robust production of both ZIKV prototype African-lineage MR766 and clinical Asian-lineage FSS13025 strains using a tetracycline (Tet)-controlled gene expression vector. Read More

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December 2018

Light Control of the Tet Gene Expression System in Mammalian Cells.

Cell Rep 2018 10;25(2):487-500.e6

Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan; Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto 606-8507, Japan; World Premier International Research Center Initiative, Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8501, Japan; The Hakubi Center for Advanced Research, Kyoto University, Kyoto 606-8302, Japan; Precursory Research for Embryonic Science and Technology, Japan Science and Technology Agency, Saitama 332-0012, Japan. Electronic address:

Gene expression and its network structure are dynamically altered in multicellular systems during morphological, functional, and pathological changes. To precisely analyze the functional roles of dynamic gene expression changes, tools that manipulate gene expression at fine spatiotemporal resolution are needed. The tetracycline (Tet)-controlled gene expression system is a reliable drug-inducible method, and it is used widely in many mammalian cultured cells and model organisms. Read More

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October 2018

Controlled re-activation of epigenetically silenced Tet promoter-driven transgene expression by targeted demethylation.

Nucleic Acids Res 2017 Sep;45(16):e147

Helmholtz Centre for Infection Research, RG Model Systems for Infection and Immunity (MSYS), Braunschweig, Germany.

Faithful expression of transgenes in cell cultures and mice is often challenged by locus dependent epigenetic silencing. We investigated silencing of Tet-controlled expression cassettes within the mouse ROSA26 locus. We observed pronounced DNA methylation of the Tet promoter concomitant with loss of expression in mES cells as well as in differentiated cells and transgenic animals. Read More

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September 2017

Controlled Osteogenic Differentiation of Mouse Mesenchymal Stem Cells by Tetracycline-Controlled Transcriptional Activation of Amelogenin.

PLoS One 2015 28;10(12):e0145677. Epub 2015 Dec 28.

Department of Fixed Prosthodontics, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan.

Regenerative dental therapies for bone tissues rely on efficient targeting of endogenous and transplanted mesenchymal stem cells (MSCs) to guide bone formation. Amelogenin is the primary component of Emdogain, which is used to regenerate periodontal defects; however, the mechanisms underlying the therapeutic effects on alveolar bone remain unclear. The tetracycline (Tet)-dependent transcriptional regulatory system is a good candidate to investigate distinct roles of genes of interest during stem cell differentiation. Read More

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Fluorescent Calcium Indicator Protein Expression in the Brain Using Tetracycline-Responsive Transgenic Mice.

Cold Spring Harb Protoc 2015 Jul 1;2015(7):689-96. Epub 2015 Jul 1.

To achieve robust long-term fluorescent calcium indicator protein (FCIP) expression in mammalian neurons in vivo, classical mouse transgenesis by pronuclear DNA injection using tetracycline (Tet)-controlled genetic switches can be deployed. This protocol describes methods for regulated expression of FCIP using Tet-responsive transgenic mice. The Tet-inducible system requires three components for inducible and reversible control of gene expression: (1) a potent transcriptional activator protein, either Tet transactivator (tTA) or reverse tTA (rtTA); (2) a minimal Tet-promoter (P(tet)) or a bidirectional Tet-promoter (P(tet)bi) to express one or more responder genes; and (3) Tet or one of its derivatives such as doxycycline (Dox) as an inducer. Read More

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Generation of transgenic fibroblasts producing doxycycline-inducible human interferon-α or erythropoietin for a bovine mammary bioreactor.

Mol Med Rep 2015 Jul 13;12(1):1137-44. Epub 2015 Mar 13.

Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 361‑763, Republic of Korea.

Interferon α (IFN-α) is a cytokine, produced predominantly in immune cells in response to pathogens, which interferes with viral replication in host cells. Another cytokine hormone, erythropoietin (EPO), is synthesized in interstitial fibroblasts of the kidney and acts as a stimulator for the production of red blood cells. Importantly, the two cytokines have been used in the treatment of certain hematological malignancies, including renal anemia. Read More

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Generation of transgenic fibroblasts expressing pancreas‑specific and doxycycline-inducible ICER Iγ for the establishment of a porcine model of human diabetes mellitus.

Mol Med Rep 2014 Aug 20;10(2):1136-42. Epub 2014 May 20.

Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 361-763, Republic of Korea.

Inducible cyclic AMP (cAMP) early repressor (ICER) Iγ acts as an endogenous inhibitor and disrupts the transcriptional regulation of cAMP response element binding protein (CREBP) responsive genes. Since the overexpression of ICER Iγ induces severe diabetes in a transgenic mouse model, with characteristics similar to human diabetes mellitus, an ICER Iγ construct containing an adjustable pancreas tissue specific promoter was utilized in the present study. Using the human insulin promoter region, a doxycycline (dox)‑inducible ICER Iγ expression system was established using the tetracycline (tet)-controlled transactivator (tTA) with a TA response element (TRE) promoter. Read More

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Inducible, tightly regulated and non-leaky neuronal gene expression in mice.

Transgenic Res 2014 Apr 9;23(2):225-33. Epub 2013 Nov 9.

Transgenic Animal Unit, School of Medical Science, University of New South Wales, Sydney, NSW, 2052, Australia.

The Tetracycline (Tet)-controlled inducible system is the most widely used reversible system for transgene expression in mice with over 500 lines created to date. Although this system has been optimized over the years, it still has limitations such as residual transgene expression when turned off, referred to as leakiness. Here, we present a series of new Tet-OFF transgenic mice based on the second generation tetracycline-responsive transactivator system. Read More

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Airway-specific inducible transgene expression using aerosolized doxycycline.

Am J Respir Cell Mol Biol 2013 Dec;49(6):1048-56

1 Center for Regenerative Medicine, and.

Tissue-specific transgene expression using tetracycline (tet)-regulated promoter/operator elements has been used to revolutionize our understanding of cellular and molecular processes. However, because most tet-regulated mouse strains use promoters of genes expressed in multiple tissues, to achieve exclusive expression in an organ of interest is often impossible. Indeed, in the extreme case, unwanted transgene expression in other organ systems causes lethality and precludes the study of the transgene in the actual organ of interest. Read More

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December 2013

Tightly regulated and homogeneous transgene expression in human adipose-derived mesenchymal stem cells by lentivirus with tet-off system.

PLoS One 2013 12;8(6):e66274. Epub 2013 Jun 12.

Pharmaceutical Research and Technology Institute, Kinki University, Higashi-Osaka, Osaka, Japan.

Genetic modification of human adipose tissue-derived multilineage progenitor cells (hADMPCs) is highly valuable for their exploitation in therapeutic applications. Here, we have developed a novel single tet-off lentiviral vector platform. This vector combines (1) a modified tetracycline (tet)-response element composite promoter, (2) a multi-cistronic strategy to express an improved version of the tet-controlled transactivator and the blasticidin resistance gene under the control of a ubiquitous promoter, and (3) acceptor sites for easy recombination cloning of the gene of interest. Read More

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January 2014

RANGE: Gene Transfer of Reversibly Controlled Polycistronic Genes.

Mol Ther Nucleic Acids 2013 Apr 9;2:e85. Epub 2013 Apr 9.

Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Heidelberg, Germany.

We developed a single vector recombinant adeno-associated viral (rAAV) expression system for spatial and reversible control of polycistronic gene expression. Our approach (i) integrates the advantages of the tetracycline (Tet)-controlled transcriptional silencer tTS(Kid) and the self-cleaving 2A peptide bridge, (ii) combines essential regulatory components as an autoregulatory loop, (iii) simplifies the gene delivery scheme, and (iv) regulates multiple genes in a synchronized manner. Controlled by an upstream Tet-responsive element (TRE), both the ubiquitous chicken β-actin promoter (CAG) and the neuron-specific synapsin-1 promoter (Syn) could regulate expression of tTS(Kid) together with two 2A-linked reporter genes. Read More

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Inducible gene expression in postmitotic neurons by an in vivo electroporation-based tetracycline system.

J Neurosci Methods 2013 Apr 25;214(2):170-6. Epub 2013 Jan 25.

Department of Developmental Biology, Graduate School of Medicine, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8670, Japan.

In vivo electroporation has been widely used to transfect foreign genes into neural progenitors and analyze the function of genes of interest in the developing nervous system. However, it has not been thoroughly examined in the conditional regulation of exogenous genes in postmitotic neurons. Here we show that the combination of in vivo electroporation and the newest version of the tetracycline (Tet)-controlled gene regulatory (Tet-On) system efficiently induced gene expression in various types of neurons in mouse embryonic and postnatal tissues. Read More

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Graded or threshold response of the tet-controlled gene expression: all depends on the concentration of the transactivator.

BMC Biotechnol 2013 Jan 22;13. Epub 2013 Jan 22.

EUFETS GmbH, Idar-Oberstein, 55743, Germany.

Background: Currently, the step-wise integration of tet-dependent transactivator and tet-responsive expression unit is considered to be the most promising tool to achieve stable tet-controlled gene expression in cell populations. However, disadvantages of this strategy for integration into primary cells led us to develop an "All-In-One" vector system, enabling simultaneous integration of both components. The effect on tet-controlled gene expression was analyzed for retroviral "All-In-One" vectors expressing the M2-transactivator either under control of a constitutive or a new type of autoregulated promoter. Read More

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January 2013

A general approach for controlling transcription and probing epigenetic mechanisms: application to the CD4 locus.

J Immunol 2013 Jan;190(2):737-47

Department of Immunobiology, Yale University Medical School, New Haven, CT 06520, USA.

Synthetic regulatory proteins such as tetracycline (tet)-controlled transcription factors are potentially useful for repression as well as ectopic activation of endogenous genes and also for probing their regulatory mechanisms, which would offer a versatile genetic tool advantageous over conventional gene targeting methods. In this study, we provide evidence supporting this concept using Cd4 as a model. CD4 is expressed in double-positive and CD4 cells but irreversibly silenced in CD8 cells. Read More

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January 2013

Conditional gene expression systems in the transgenic rat brain.

BMC Biol 2012 Sep 3;10:77. Epub 2012 Sep 3.

Department of Molecular Biology, Central Institute of Mental Health and Heidelberg University, Medical Faculty Mannheim, J5, 68159 Mannheim, Germany.

Background: Turning gene expression on and off at will is one of the most powerful tools for the study of gene function in vivo. While several conditional systems were successful in invertebrates, in mice the Cre/loxP recombination system and the tet-controlled transcription activation system are predominant. Both expression systems allow for spatial and temporal control of gene activities, and, in the case of tet regulation, even for the reversible activation/inactivation of gene expression. Read More

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September 2012

Doxycycline-regulated growth hormone gene expression system for swine.

Genet Mol Res 2012 Aug 29;11(3):2946-57. Epub 2012 Aug 29.

Department of Animal Breeding & Genetics, College of Animal Science & Technology, Nanjing Agricultural University, Nanjing, China.

Although growth hormone gene transgenic animals are much larger than normal animals, they manifest differences that have adverse effects on survival due to overexpression of growth hormone. We developed a stable pig embryonic fibroblast cell line expressing pig growth hormone (pGH) using the Tet-On system, with which we can conditionally manipulate expression of pGH in vivo. Inducible expression of pGH was achieved by combining reserve Tet-controlled transcriptional activator and tetracycline-responsive element in a single plasmid. Read More

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Promoter strength driving TetR determines the regulatory properties of Tet-controlled expression systems.

PLoS One 2012 27;7(7):e41620. Epub 2012 Jul 27.

Lehrstuhl für Mikrobiologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.

Bacteria frequently rely on transcription repressors and activators to alter gene expression patterns in response to changes in the surrounding environment. Tet repressor (TetR) is a paradigm transcription factor that senses the environmental state by binding small molecule effectors, the tetracyclines. However, recently isolated peptides that act as inducers of TetR after having been fused to the C-terminus of a carrier protein, suggest that TetR can also regulate gene expression in a signal-transduction pathway. Read More

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Protein expression can be monitored in yeast by peptide-mediated induction of TetR-controlled gene expression.

J Biotechnol 2012 Oct 20;161(3):265-8. Epub 2012 Jun 20.

Lehrstuhl für Mikrobiologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstrasse 5, 91058 Erlangen, Germany.

The rapidly increasing number of completed genome sequences urgently calls for convenient and efficient methods for analysis of gene function and expression. TetR-inducing peptides (TIP) can induce reporter gene expression controlled by Tet repressor (TetR) when fused to a protein of choice which makes them a highly valuable tool for monitoring expression in vivo. However, TIP functionality has only been demonstrated in bacteria so far. Read More

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October 2012

First inducible transgene expression in porcine large animal models.

FASEB J 2012 Mar 1;26(3):1086-99. Epub 2011 Dec 1.

Ludwig-Maximilians-Universitat Munchen, Munich, Germany.

The purpose of this study was to establish inducible transgene expression in pigs, a model organism with great promise for experimental physiology and translational medicine, using the binary tet-on system. This expression system is activated by doxycycline (dox) via the tet-controlled transactivator (TA). Binding of TA to the transactivator response element (TRE) results in transcription of downstream genes. Read More

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Inducible microRNA-mediated knockdown of the endogenous human lamin A/C gene.

Authors:
Ina Weidenfeld

Methods Mol Biol 2012 ;815:289-305

Molecular, Cellular, and Developmental Biology, University of Colorado at Boulder, Boulder, CO 80309, USA.

RNA interference (RNAi) enables the suppression, and hence the functional analysis, of individual genes. The use of the tetracycline (tet)-controlled transcription activation system for RNAi has become a valuable tool for conditional gene inactivation both in vitro and in vivo. Here, the generation of a conditional RNAi cell line for microRNA (miRNA)-mediated downregulation of the endogenous lamin A/C gene is described. Read More

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Development of a BAC vector for integration-independent and tight regulation of transgenes in rodents via the Tet system.

Transgenic Res 2011 Jun 18;20(3):709-20. Epub 2010 Jul 18.

Department of Molecular Biology, Central Institute of Metal Health and Heidelberg University, Medical Faculty Mannheim, J5, 68159, Mannheim, Germany.

The establishment of functional transgenic mouse lines is often limited by problems caused by integration site effects on the expression construct. Similarly, tetracycline (Tet) controlled transcription units most commonly used for conditional transgene expression in mice are strongly influenced by their genomic surrounding. Using bacterial artificial chromosome (BAC) technology in constitutive expression systems, it has been shown that integration site effects resulting in unwanted expression patterns can be largely eliminated. Read More

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Quantitative analysis of conditional gene inactivation using rationally designed, tetracycline-controlled miRNAs.

Nucleic Acids Res 2010 Sep 17;38(17):e168. Epub 2010 Jul 17.

Central Institute of Mental Health, Department of Molecular Biology, D-68159 Mannheim, Germany.

The combination of RNA interference (RNAi) with the tetracycline-controlled transcription activation (tet) system promises to become a powerful method for conditional gene inactivation in cultured cells and in whole organisms. Here, we tested critical sequence elements that originated from miRNA mR-30 for optimal efficiency of RNAi-based gene knockdown in mammalian cells. Rationally designed miRNAs, expressed conditionally via the tet system, led to an efficient knockdown of the expression of both reporter genes and the endogenous mitotic spindle protein TPX2 in HeLa cells. Read More

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September 2010

Optogenetic Dissection of Neuronal Circuits in Zebrafish using Viral Gene Transfer and the Tet System.

Front Neural Circuits 2009 11;3:21. Epub 2009 Dec 11.

Friedrich Miescher Institute for Biomedical Research, Novartis Research Foundation Basel, Switzerland.

The conditional expression of transgenes at high levels in sparse and specific populations of neurons is important for high-resolution optogenetic analyses of neuronal circuits. We explored two complementary methods, viral gene delivery and the iTet-Off system, to express transgenes in the brain of zebrafish. High-level gene expression in neurons was achieved by Sindbis and Rabies viruses. Read More

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Improved applications of the tetracycline-regulated gene depletion system.

Biosci Trends 2009 Oct;3(5):161-7

Department of Integrated Genetics, National Institute of Genetics, Mishima, Shizuoka, Japan.

Tightly controlled expression of transgenes in mammalian cells is an important tool for biological research, drug discovery, and future genetic therapies. The tetracycline-regulated gene depletion (Tet-Off) system has been widely used to control gene activities in mammalian cells, because it allows strict regulation of transgenes but no pleiotropic effects of prokaryotic regulatory proteins. However, the Tet-Off system is not compatible with every cell type and this is the main remaining obstacle left for this system. Read More

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October 2009

Select overexpression of homer1a in dorsal hippocampus impairs spatial working memory.

Front Neurosci 2007 Nov 15;1(1):97-110. Epub 2007 Oct 15.

Department of Cell Physiology, Max-Planck Institute for Medical Research, Heidelberg Germany.

Long Homer proteins forge assemblies of signaling components involved in glutamate receptor signaling in postsynaptic excitatory neurons, including those underlying synaptic transmission and plasticity. The short immediate-early gene (IEG) Homer1a can dynamically uncouple these physical associations by functional competition with long Homer isoforms. To examine the consequences of Homer1a-mediated "uncoupling" for synaptic plasticity and behavior, we generated forebrain-specific tetracycline (tet) controlled expression of Venus-tagged Homer1a (H1aV) in mice. Read More

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November 2007