Keratin promoter based gene manipulation in the murine conducting airway

Stephen P. Malkoski, Timothy G. Cleaver, Shi Long Lu, Jessyka G. Lighthall, Xiao Jing Wang

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Systems capable of targeting genetic manipulations to keratin-positive airway basal cells are more poorly developed than systems targeting other airway epithelial cell populations and this has likely hindered development of animal models of diseases such as lung squamous cell carcinoma. Although keratin promoter driven-Cre recombinase constructs are potentially useful for targeting these cells, these constructs have substantially higher activity in the skin and oral epithelium than in the airways. We developed a method for delivering RU486, the conditional activator of Cre recombinase progesterone receptor (CrePR) fusion proteins to the lung and then examined the activity of three keratin-driven CrePR constructs in the conducting airways. We also developed a technique for survival bronchioalveolar lavage on non-ventilated animals to examine the effects of the acetone/oil vehicle required to deliver RU486 to the lung. K5CrePR1 and K14CrePR1 constructs differ only in the keratin promoter used to target CrePR1 expression while K5Cre*PR contains a truncated progester-one receptor designed to reduce RU486-independent Cre activity. While all three constructs demonstrate RU486-inducible Cre activity in the conducting airways, both construct activity and tightness of regulation vary considerably. K5Cre*PR is the most tightly regulated Cre driver making it ideal for targeting somatic mutations to the airway epithelia while K5CrePR1 and K14CrePR1 may be better suited to studying diseases of the conducting airways where gene targeting of keratin expressing cells and their derivatives is desired.

Original languageEnglish (US)
Pages (from-to)68-79
Number of pages12
JournalInternational Journal of Biological Sciences
Volume6
Issue number1
DOIs
StatePublished - Jan 1 2010

Fingerprint

keratin
Keratins
targeting
promoter regions
gene
mice
Genes
lungs
Progesterone Receptors
genes
Lung
epithelium
Epithelium
animal disease models
Animal Disease Models
somatic mutation
Gene Targeting
animal
squamous cell carcinoma
Bronchoalveolar Lavage

All Science Journal Classification (ASJC) codes

  • Ecology, Evolution, Behavior and Systematics
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Developmental Biology
  • Cell Biology

Cite this

Malkoski, Stephen P. ; Cleaver, Timothy G. ; Lu, Shi Long ; Lighthall, Jessyka G. ; Wang, Xiao Jing. / Keratin promoter based gene manipulation in the murine conducting airway. In: International Journal of Biological Sciences. 2010 ; Vol. 6, No. 1. pp. 68-79.
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Keratin promoter based gene manipulation in the murine conducting airway. / Malkoski, Stephen P.; Cleaver, Timothy G.; Lu, Shi Long; Lighthall, Jessyka G.; Wang, Xiao Jing.

In: International Journal of Biological Sciences, Vol. 6, No. 1, 01.01.2010, p. 68-79.

Research output: Contribution to journalArticle

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