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. 2007 May;36(5):515-9.
doi: 10.1165/rcmb.2006-0475RC. Epub 2007 Jan 25.

A transgenic FOXJ1-Cre system for gene inactivation in ciliated epithelial cells (VSports在线直播)

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A transgenic FOXJ1-Cre system for gene inactivation in ciliated epithelial cells

Yong Zhang et al. Am J Respir Cell Mol Biol. 2007 May.

"VSports在线直播" Abstract

Ciliated airway epithelial cells are critical for mucosal barrier function, including host defense against pathogens. This cell population is often the primary target and thereby the first line of defense against many common respiratory viruses. It is also the precursor for mucous cells and thereby promotes mucociliary clearance of infectious and other noxious agents. Cells with motile cilia in other organs (e. g. , brain and reproductive organs) may also have roles in development and reproduction VSports手机版. However, definitive proof of ciliated cell function is hampered by the lack of strategies to specifically target this cell population for loss of function in vivo. To this end, cell type-specific gene promoters have been combined with the Cre/LoxP system to disrupt genes in airway and alveolar epithelial cell populations expressing surfactant protein C (SP-C) or Clara cell secretory protein (CCSP). By contrast, an analogous system to disrupt gene function in ciliated airway epithelial cells was still needed. Here we report the generation and analysis of mouse lines with a FOXJ1 promoter driving the Cre recombinase and show that this system mediates genomic recombination specifically in ciliated cells. The pattern of recombination recapitulates endogenous FOXJ1 promoter function, being restricted to ciliated cells present in pulmonary airways as well as choroid plexus, ependyma, oviduct, and testis. This transgenic mouse system thereby offers a new strategy for specific knockouts of genes in ciliated cells. It should prove extremely useful for defining ciliated cell function in airway mucosal immunity as well as development and reproduction. .

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Figures (V体育ios版)

<b>Figure 1.</b>
Figure 1.
Tissue localization of Cre expression in FOXJ1-Cre transgenic mice. (a) Schematic representation of DNA construct used to generate the FOXJ1-Cre transgenic mouse line. (b) Analysis of Cre transgene expression in indicated tissues isolated from FOXJ1-Cre transgenic mice and then subjected to real-time PCR for Cre mRNA levels. Values represent mean levels for two to three mice from each of three founder lines (F14, F24, and F26) and are normalized to Gapdh mRNA levels in each sample.
<b>Figure 2.</b>
Figure 2.
Cellular localization of Cre-mediated recombination in FOXJ1-Cre/ROSA26R double-transgenic mice. (a) Schematic representation of breeding cross between FOXJ1-Cre and ROSA transgenic mice. (b) Tracheal and lung cryostat sections from FOXJ1-Cre/ROSA double-transgenic and Cre-negative ROSA control mice were stained with X-gal and counterstained with eosin. Results are representative of three FOXJ1-Cre/ROSA founder lines. Insets represent higher magnification views. Bars = 40 μm. (c) Tracheal cryostat sections from FOXJ1-Cre/ROSA mice and Cre-negative ROSA control mice were immunostained with anti–β-Gal antibody detected with Cy3-labeled secondary antibody (red fluorescence) and anti–β-Tubulin mAb detected with FITC-labeled secondary antibody (green fluorescence). Nuclear DNA was detected with DAPI (blue fluorescence). Bar = 20 μm. (d) mTEC cultures from FOXJ1-Cre/ROSA mice were immunostained using the same antibodies as in c. Cellular β-Gal staining was not detected in Cre-negative ROSA control mice (data not shown). Bar = 20 μm.
<b>Figure 3.</b>
Figure 3.
Tissue localization of Cre-mediated recombination in FOXJ1-Cre/ROSA26R mice. (a) Tissue cryostat sections from testis, oviduct, ependyma, and choroid plexus of FOXJ1-Cre/ROSA double-transgenic mice were stained with X-Gal and counterstained with eosin. Insets represent higher magnification views. (b) Tissue cryostat sections from heart, intestine, liver, and kidney of mice in a were stained with X-Gal and counterstained with eosin. Bars = 40 μm.

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