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Article: Regulated expression of the human CFTR gene in epithelial cells

TitleRegulated expression of the human CFTR gene in epithelial cells
Authors
KeywordsCFTR
Epithelia
Gene expression
Inducible
Transgenic mice
Issue Date2001
PublisherNature Publishing Group.
Citation
Molecular Therapy, 2001, v. 3 n. 5 I, p. 723-733 How to Cite?
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.
Persistent Identifierhttp://hdl.handle.net/10722/44366
ISSN
2014 Impact Factor: 6.227
2014 SCImago Journal Rankings: 2.687
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorYe, Len_HK
dc.contributor.authorChan, Sen_HK
dc.contributor.authorChow, YHen_HK
dc.contributor.authorTsui, LCen_HK
dc.contributor.authorHu, Jen_HK
dc.date.accessioned2007-09-12T03:52:11Z-
dc.date.available2007-09-12T03:52:11Z-
dc.date.issued2001en_HK
dc.identifier.citationMolecular Therapy, 2001, v. 3 n. 5 I, p. 723-733en_HK
dc.identifier.issn1525-0016en_HK
dc.identifier.urihttp://hdl.handle.net/10722/44366-
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.en_HK
dc.languageengen_HK
dc.publisherNature Publishing Group.en_HK
dc.relation.ispartofMolecular Therapyen_HK
dc.subjectCFTRen_HK
dc.subjectEpitheliaen_HK
dc.subjectGene expressionen_HK
dc.subjectInducibleen_HK
dc.subjectTransgenic miceen_HK
dc.subject.meshGene expressionen_HK
dc.subject.meshEpitheliaen_HK
dc.subject.meshCftren_HK
dc.subject.meshTransgenic miceen_HK
dc.subject.meshInducibleen_HK
dc.titleRegulated expression of the human CFTR gene in epithelial cellsen_HK
dc.typeArticleen_HK
dc.identifier.emailTsui, LC: tsuilc@hkucc.hku.hken_HK
dc.identifier.authorityTsui, LC=rp00058en_HK
dc.description.naturelink_to_OA_fulltexten_HK
dc.identifier.doi10.1006/mthe.2001.0314en_HK
dc.identifier.pmid11356077en_HK
dc.identifier.scopuseid_2-s2.0-0034981141en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0034981141&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume3en_HK
dc.identifier.issue5 Ien_HK
dc.identifier.spage723en_HK
dc.identifier.epage733en_HK
dc.identifier.isiWOS:000168924400011-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridYe, L=35504324300en_HK
dc.identifier.scopusauthoridChan, S=36901167700en_HK
dc.identifier.scopusauthoridChow, YH=7202906132en_HK
dc.identifier.scopusauthoridTsui, LC=7102754167en_HK
dc.identifier.scopusauthoridHu, J=7406420553en_HK

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