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Article: Regulated expression of the human CFTR gene in epithelial cells
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TitleRegulated expression of the human CFTR gene in epithelial cells
 
AuthorsYe, L2 1
Chan, S2 1
Chow, YH2 1
Tsui, LC2 1
Hu, J2 1
 
KeywordsCFTR
Epithelia
Gene expression
Inducible
Transgenic mice
 
Issue Date2001
 
PublisherNature Publishing Group.
 
CitationMolecular Therapy, 2001, v. 3 n. 5 I, p. 723-733 [How to Cite?]
DOI: http://dx.doi.org/10.1006/mthe.2001.0314
 
AbstractWe developed an epithelium-specific, inducible cystic fibrosis transmembrane conductance regulator (CFTR) expression system. In this system we used a human cytokeratin 18 expression cassette to drive epithelium-specific expression of the reverse tetracycline transactivator (rtTA), which turns on CFTR expression from a Tet-inducible promoter in the presence of doxycycline. CFTR expression was monitored by reverse-transcription polymerase chain reaction, immunostaining, and Western blotting. We confirmed that protein expression was dose-dependent in double stable transfected cell lines, with no detectable protein in the absence of doxycycline. However, low levels of CFTR mRNA could be detected in the uninduced state. When clones capable of inducing high levels of CFTR expression were analyzed, we observed a decrease in cell proliferation, consistent with reports in other cell lines (NIH3T3 and BTS). We generated transgenic mice expressing rtTA from the K18 expression cassette and demonstrated that the system retained its tissue specificity for lacZ reporter expression in vivo. When mice were induced with doxycycline, high levels of expression were found in the trachea, upper bronchi, and submucosal glands. Therefore, this inducible system can improve our understanding of the role of CFTR in the lung and should help in the design of safe and effective CF therapies.
 
ISSN1525-0016
2013 Impact Factor: 6.425
 
DOIhttp://dx.doi.org/10.1006/mthe.2001.0314
 
ISI Accession Number IDWOS:000168924400011
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorYe, L
 
dc.contributor.authorChan, S
 
dc.contributor.authorChow, YH
 
dc.contributor.authorTsui, LC
 
dc.contributor.authorHu, J
 
dc.date.accessioned2007-09-12T03:52:11Z
 
dc.date.available2007-09-12T03:52:11Z
 
dc.date.issued2001
 
dc.description.abstractWe developed an epithelium-specific, inducible cystic fibrosis transmembrane conductance regulator (CFTR) expression system. In this system we used a human cytokeratin 18 expression cassette to drive epithelium-specific expression of the reverse tetracycline transactivator (rtTA), which turns on CFTR expression from a Tet-inducible promoter in the presence of doxycycline. CFTR expression was monitored by reverse-transcription polymerase chain reaction, immunostaining, and Western blotting. We confirmed that protein expression was dose-dependent in double stable transfected cell lines, with no detectable protein in the absence of doxycycline. However, low levels of CFTR mRNA could be detected in the uninduced state. When clones capable of inducing high levels of CFTR expression were analyzed, we observed a decrease in cell proliferation, consistent with reports in other cell lines (NIH3T3 and BTS). We generated transgenic mice expressing rtTA from the K18 expression cassette and demonstrated that the system retained its tissue specificity for lacZ reporter expression in vivo. When mice were induced with doxycycline, high levels of expression were found in the trachea, upper bronchi, and submucosal glands. Therefore, this inducible system can improve our understanding of the role of CFTR in the lung and should help in the design of safe and effective CF therapies.
 
dc.description.naturelink_to_OA_fulltext
 
dc.identifier.citationMolecular Therapy, 2001, v. 3 n. 5 I, p. 723-733 [How to Cite?]
DOI: http://dx.doi.org/10.1006/mthe.2001.0314
 
dc.identifier.doihttp://dx.doi.org/10.1006/mthe.2001.0314
 
dc.identifier.epage733
 
dc.identifier.isiWOS:000168924400011
 
dc.identifier.issn1525-0016
2013 Impact Factor: 6.425
 
dc.identifier.issue5 I
 
dc.identifier.pmid11356077
 
dc.identifier.scopuseid_2-s2.0-0034981141
 
dc.identifier.spage723
 
dc.identifier.urihttp://hdl.handle.net/10722/44366
 
dc.identifier.volume3
 
dc.languageeng
 
dc.publisherNature Publishing Group.
 
dc.publisher.placeUnited Kingdom
 
dc.relation.ispartofMolecular Therapy
 
dc.relation.referencesReferences in Scopus
 
dc.subject.meshGene expression
 
dc.subject.meshEpithelia
 
dc.subject.meshCftr
 
dc.subject.meshTransgenic mice
 
dc.subject.meshInducible
 
dc.subjectCFTR
 
dc.subjectEpithelia
 
dc.subjectGene expression
 
dc.subjectInducible
 
dc.subjectTransgenic mice
 
dc.titleRegulated expression of the human CFTR gene in epithelial cells
 
dc.typeArticle
 
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<contributor.author>Hu, J</contributor.author>
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Author Affiliations
  1. University of Toronto
  2. Hospital for Sick Children University of Toronto