Efficiency and Specificity of Gene Deletion in Lung Epithelial Doxycycline-Inducible Cre Mice.
Am J Respir Cell Mol Biol
; 57(2): 248-257, 2017 08.
Article
em En
| MEDLINE
| ID: mdl-28287822
ABSTRACT
The transgenic mouse strains surfactant protein C-reverse tetracycline transactivator (SP-C-rtTA), club cell secretory protein (CCSP)-rtTA, and tetracycline operator (TetO)-Cre have been invaluable for spatiotemporally regulating gene deletion in the pulmonary epithelium. In this study, we measured the efficiency and specificity of gene deletion that can be achieved in these mice using the Rosa26-eYFP reporter. Triple-transgenic mice (tTg or rtTA/TetO-Cre/Rosa-eYFP) were bred and treated with various doxycycline (dox) regimens to induce gene deletion, which was then quantified in various cell populations by flow cytometry. In these crosses, we found that the TetO-Cre transgene must be transmitted through the female parent to avoid germline gene deletion. With dox exposure during lung development, SP-C-tTg mice deleted in â¼65-75% of alveolar epithelial type II (ATII) cells, but in only â¼45-50% of the integrin ß4+ population, which consisted of club cells and distal lung progenitor cells. In contrast, CCSP-tTg mice deleted in â¼50% of ATII cells and â¼80% of integrin ß4+ cells. Upon dox treatment of adults, deletion in ATII cells and integrin ß4+ cells in SP-C-tTg mice dropped significantly to â¼20% and â¼6%, respectively, whereas CCSP-tTg mice deleted in â¼57% of ATII and â¼40% of integrin ß4+ cells. Interestingly, untreated CCSP-tTg mice also deleted in â¼40% of integrin ß4+ cells, indicating significant leakiness of CCSP-tTg in ß4+ cells. In all mouse groups, minimal deletion occurred in mouse tracheal epithelial cells or in mesenchymal or hematopoietic cells. These data provide the first quantitative, side-by-side comparison of the deletion efficiency for these widely used transgenic mouse strains.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Traqueia
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Camundongos Transgênicos
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Deleção de Genes
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Transgenes
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Integrases
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Técnicas de Inativação de Genes
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Células Epiteliais Alveolares
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Pulmão
Limite:
Animals
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article