Use of the Tetracycline-controlled Transcriptional Silencer (tTS) to Eliminate Transgene Leak in Inducible Overexpression Transgenic Mice

Zhu, Zhou; Ma, Bing; Homer, Robert J.; Zheng, Tao; Elias, Jack A.

doi:10.1074/jbc.m101512200

Cited by 128 publications

(84 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Ideally, age-specific TGF-␤ 1 overexpression, as well as selective modulation of lung epithelial cell T␤R expression by transgenic technologies, will help to address these questions in the future. At present, only the selective overexpression of soluble secreted proteins in the lung is a feasible approach to analyze the morphological effects of growth factors or cytokines (35). The presented findings, nevertheless, suggest an essential role for the TGF-␤ system in the response of lung epithelial cells to chronic hypoxia at different developmental stages.…”

Section: Discussionmentioning

confidence: 77%

Regulation of TGF-β ligand and receptor expression in neonatal rat lungs exposed to chronic hypoxia

Vicencio¹,

Eickelberg

Stankewich

et al. 2002

Journal of Applied Physiology

View full text Add to dashboard Cite

Regulation of TGF-␤ ligand and receptor expression in neonatal rat lungs exposed to chronic hypoxia. J Appl Physiol 93: 1123-1130. First published May 3, 2002 10.1152/japplphysiol.00031.2002.-Long-term effects of hypoxia are largely due to its modulatory effects on proliferation and differentiation of epithelial and endothelial cells, processes also regulated by the transforming growth factor (TGF)-␤ system. We investigated the effects of hypoxia on the TGF-␤ system in rat lungs from different developmental stages. Sprague-Dawley rats were exposed to 9.5% oxygen during either the first 2 wk of life or adulthood. Analysis revealed an arrest of alveolarization in hypoxic postnatal day 14 rats. Bioactive TGF-␤ levels in bronchoalveolar lavage fluid were increased in these animals, and Western blot analysis revealed upregulation of TGF-␤ receptor (T␤R) I and II. None of these changes was observed in hypoxic adults. Hypoxia did, however, lead to decreased expression of T␤RIII in both postnatal day 14 and adult rats. Immunohistochemical analysis localized T␤RI-III predominantly to bronchiolar and alveolar epithelium; these patterns did not change with hypoxia. Thus we observed changes in TGF-␤ activity and T␤R isotype expression in rat lung that parallel the arrest in alveolarization seen with chronic hypoxia in early development. These alterations may partly explain the morphological changes observed in hypoxia.transforming growth factor-␤; lung development; alveolarization; transforming growth factor-␤ receptor; betaglycan POSTNATAL LUNG DEVELOPMENT involves the formation of alveoli from existing but immature respiratory saccules by means of septation, maturation of epithelial and mesenchymal units, and development of the alveolar capillary system (4, 21). In rats, alveolar septation begins at postnatal days (P) 3-4 and continues throughout the second week of postnatal development. Maturation of newly formed units occurs in parallel and continues throughout the fourth week of postnatal lung development. These processes are tightly regulated by direct interactions between epithelial and mesenchymal cells (5).Several intrinsic and extrinsic factors have been identified as modifiers of postnatal lung growth. For example, hypoxia over prolonged periods can affect lung growth (27). There has been some controversy, however, regarding the overall effects of hypoxia on the postnatal development of the lung (3, 22). Whether such issues are related to age or stage of development at the time of exposure is not well defined, and possible mediators of the effects of hypoxia on lung morphology remain essentially unexplored.Whereas the molecular mechanisms underlying the effects of hypoxia on lung growth remain to be delineated, significant advances have been made in our understanding of the mediators controlling prenatal lung morphogenesis and branching. Members of the transforming growth factor (TGF)-␤ family have emerged as crucial factors controlling the formation, patterning, and maturation of lung tissue (13, 24). TGF-␤ bel...

show abstract

Section: Discussionmentioning

confidence: 77%

Regulation of TGF-β ligand and receptor expression in neonatal rat lungs exposed to chronic hypoxia

Vicencio¹,

Eickelberg

Stankewich

et al. 2002

Journal of Applied Physiology

View full text Add to dashboard Cite

show abstract

“…The final plasmid ((tet-O) 7 -FLAG-cIKK2-BGH.poly(A)) was verified by sequencing. To prevent basal leakiness, we used a construct expressing a tetracycline-controlled transcriptional silencer (tTS) under control of the CC10 promoter (CC10-tTS-hGH-poly(A)) (10). The (tet-O) 7 -FLAG-cIKK2 microinjection fragment was excised from the plasmid as a 3.3-kb XmnI-AscI fragment.…”

Section: Transgenic Mouse Modelsmentioning

confidence: 99%

“…Farmer, BD Clontech) was coinjected with (tet-O) 7 -FLAG-cIKK2 to generate double transgenic mice. Unbound tTS interacts with tet-O sites and functions as a transcriptional repressor; however, binding of dox to tTS results in dissociation from DNA, allowing rtTA binding and promoter activation (10,17,18). Double transgenic mice were bred with transgenic mice expressing rtTA under the control of the rat CC10 promoter (obtained from Dr. J.…”

Section: Construction Of Transgenic Mice With Inducible Activation Ofmentioning

confidence: 99%

Airway Epithelium Controls Lung Inflammation and Injury through the NF-κB Pathway

Cheng

Han

Chen

et al. 2007

The Journal of Immunology

162

161

View full text Add to dashboard Cite

Although airway epithelial cells provide important barrier and host defense functions, a crucial role for these cells in development of acute lung inflammation and injury has not been elucidated. We investigated whether NF-κB pathway signaling in airway epithelium could decisively impact inflammatory phenotypes in the lungs by using a tetracycline-inducible system to achieve selective NF-κB activation or inhibition in vivo. In transgenic mice that express a constitutively active form of IκB kinase 2 under control of the epithelial-specific CC10 promoter, treatment with doxycycline induced NF-κB activation with consequent production of a variety of proinflammatory cytokines, high-protein pulmonary edema, and neutrophilic lung inflammation. Continued treatment with doxycycline caused progressive lung injury and hypoxemia with a high mortality rate. In contrast, inducible expression of a dominant inhibitor of NF-κB in airway epithelium prevented lung inflammation and injury resulting from expression of constitutively active form of IκB kinase 2 or Escherichia coli LPS delivered directly to the airways or systemically via an osmotic pump implanted in the peritoneal cavity. Our findings indicate that the NF-κB pathway in airway epithelial cells is critical for generation of lung inflammation and injury in response to local and systemic stimuli; therefore, targeting inflammatory pathways in airway epithelium could prove to be an effective therapeutic strategy for inflammatory lung diseases.

show abstract

“…The first batch of transgenic fish was generated three decades ago through microinjection of plasmids into fertilized eggs (Zhu et al, 1985). Since then, many commercial fish species such as carp, tilapia and salmonids have been successfully used for transgenesis (Martinez et al, 1996;Cook et al, 2000;Sun et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

“…Although the Tet-controlled systems have been widely utilized in cultured cells from plants to mammals and in organisms from Drosophila to mice (Zhu et al, 2001; A: the pTRE-DKK1 plasmids were microinjected into wild-type (WT) and F 3 transgenic embryos. Injected embryos at 1 hpf were treated with or without Dox (10 mg/mL) for 3 h. Developing embryos were fixed and processed for whole-mount in suit hybridization (WISH) with marker genes as indicated on the top.…”

Section: Discussionmentioning

confidence: 99%

Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator

Gu¹,

Yang²,

He³

et al. 2013

Journal of Genetics and Genomics

View full text Add to dashboard Cite

Use of the Tetracycline-controlled Transcriptional Silencer (tTS) to Eliminate Transgene Leak in Inducible Overexpression Transgenic Mice

Cited by 128 publications

References 33 publications

Regulation of TGF-β ligand and receptor expression in neonatal rat lungs exposed to chronic hypoxia

Regulation of TGF-β ligand and receptor expression in neonatal rat lungs exposed to chronic hypoxia

Airway Epithelium Controls Lung Inflammation and Injury through the NF-κB Pathway

Generation and Characterization of a Transgenic Zebrafish Expressing the Reverse Tetracycline Transactivator

Contact Info

Product

Resources

About