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Airway-specific inducible transgene expression using aerosolized doxycycline.

Publication ,  Journal Article
Tata, PR; Pardo-Saganta, A; Prabhu, M; Vinarsky, V; Law, BM; Fontaine, BA; Tager, AM; Rajagopal, J
Published in: Am J Respir Cell Mol Biol
December 2013

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. Here, we describe a novel approach to activating tet-inducible transgene expression solely in the airway by administering aerosolized doxycycline. By optimizing the dose and duration of aerosolized doxycycline exposure in mice possessing a ubiquitously expressed Rosa26 promoter-driven reverse tet-controlled transcriptional activator (rtTA) element, we induce transgene expression exclusively in the airways. We detect no changes in the cellular composition or proliferative behavior of airway cells. We used this newly developed method to achieve airway basal stem cell-specific transgene expression using a cytokeratin 5 (also known as keratin 5)-driven rtTA driver line to induce Notch pathway activation. We observed a more robust mucous metaplasia phenotype than in mice receiving doxycycline systemically. In addition, unwanted phenotypes outside of the lung that were evident when doxycycline was received systemically were now absent. Thus, our approach allows for rapid and efficient airway-specific transgene expression. After the careful strain by strain titration of the dose and timing of doxycycline inhalation, a suite of preexisting transgenic mice can now be used to study airway biology specifically in cases where transient transgene expression is sufficient to induce a phenotype.

Duke Scholars

Published In

Am J Respir Cell Mol Biol

DOI

EISSN

1535-4989

Publication Date

December 2013

Volume

49

Issue

6

Start / End Page

1048 / 1056

Location

United States

Related Subject Headings

  • Transgenes
  • Trans-Activators
  • Tetracycline
  • Signal Transduction
  • Respiratory System
  • Respiratory System
  • Receptors, Notch
  • RNA, Untranslated
  • Promoter Regions, Genetic
  • Phenotype
 

Citation

APA
Chicago
ICMJE
MLA
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Tata, P. R., Pardo-Saganta, A., Prabhu, M., Vinarsky, V., Law, B. M., Fontaine, B. A., … Rajagopal, J. (2013). Airway-specific inducible transgene expression using aerosolized doxycycline. Am J Respir Cell Mol Biol, 49(6), 1048–1056. https://doi.org/10.1165/rcmb.2012-0412OC
Tata, Purushothama Rao, Ana Pardo-Saganta, Mythili Prabhu, Vladimir Vinarsky, Brandon M. Law, Benjamin A. Fontaine, Andrew M. Tager, and Jayaraj Rajagopal. “Airway-specific inducible transgene expression using aerosolized doxycycline.Am J Respir Cell Mol Biol 49, no. 6 (December 2013): 1048–56. https://doi.org/10.1165/rcmb.2012-0412OC.
Tata PR, Pardo-Saganta A, Prabhu M, Vinarsky V, Law BM, Fontaine BA, et al. Airway-specific inducible transgene expression using aerosolized doxycycline. Am J Respir Cell Mol Biol. 2013 Dec;49(6):1048–56.
Tata, Purushothama Rao, et al. “Airway-specific inducible transgene expression using aerosolized doxycycline.Am J Respir Cell Mol Biol, vol. 49, no. 6, Dec. 2013, pp. 1048–56. Pubmed, doi:10.1165/rcmb.2012-0412OC.
Tata PR, Pardo-Saganta A, Prabhu M, Vinarsky V, Law BM, Fontaine BA, Tager AM, Rajagopal J. Airway-specific inducible transgene expression using aerosolized doxycycline. Am J Respir Cell Mol Biol. 2013 Dec;49(6):1048–1056.

Published In

Am J Respir Cell Mol Biol

DOI

EISSN

1535-4989

Publication Date

December 2013

Volume

49

Issue

6

Start / End Page

1048 / 1056

Location

United States

Related Subject Headings

  • Transgenes
  • Trans-Activators
  • Tetracycline
  • Signal Transduction
  • Respiratory System
  • Respiratory System
  • Receptors, Notch
  • RNA, Untranslated
  • Promoter Regions, Genetic
  • Phenotype