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Cytokeratin 19 promoter directs the expression of Cre recombinase in various epithelia of transgenic mice

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ABSTRACT

Cytokeratin 19 (K19) is expressed in various differentiated cells, including gastric, intestinal and bronchial epithelial cells, and liver duct cells. Here, we generated a transgenic mouse line, K19-Cre, in which the expression of Cre recombinase was controlled by the promoter of K19. To test the tissue distribution and excision activity of Cre recombinase, K19-Cre transgenic mice were bred with Rosa26 reporter strain and a mouse strain that carries PTEN conditional alleles (PTENLoxp/Loxp). At mRNA level, Cre was strongly expressed in the stomach, lung and intestine, while in stomach, lung, and liver at protein level. The immunoreactivity to Cre was strongly observed the cytoplasm of gastric, bronchial and intestinal epithelial cells. Cre activity was detectable in gastric, bronchial and intestinal epithelial cells, according to LacZ staining. In K19-Cre/PTEN Loxp/Loxp mice, PTEN was abrogated in stomach, intestine, lung, liver and breast, the former two of which were verified by in situ PCR. There appeared breast cancer with PTEN loss. These data suggest that K19 promoter may be a useful tool to study the pathophysiological functions of cytokeratin 19-positive cells, especially gastrointestinal epithelial cells. Cell specificity of neoplasia is not completely attributable to the cell-specific expression of oncogenes and cell-specific loss of tumor suppressor genes.

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Cre activity in various tissues of K19-Cre transgeneic miceAfter mated with Rosa26-lacZ mouse A., Cre activity in K19-Cre/Rosa transgenic mice was visualized in lung, stomach and intestine by lacZ staining B. Wild-type C57 mouse (WT) was employed as a negative control and the target knockout mice of B6.Cg-Dkk 3tm1, B6.Cg-Becn1tm1 pvillin-Cre/Rosa as a positive control due to the existence of endogenous LaZ gene. Note: PC, positive control, tail DNA of pvillin- Cre mouse or Rosa26-lacZ; NC, negative control, no DNA template.
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Figure 3: Cre activity in various tissues of K19-Cre transgeneic miceAfter mated with Rosa26-lacZ mouse A., Cre activity in K19-Cre/Rosa transgenic mice was visualized in lung, stomach and intestine by lacZ staining B. Wild-type C57 mouse (WT) was employed as a negative control and the target knockout mice of B6.Cg-Dkk 3tm1, B6.Cg-Becn1tm1 pvillin-Cre/Rosa as a positive control due to the existence of endogenous LaZ gene. Note: PC, positive control, tail DNA of pvillin- Cre mouse or Rosa26-lacZ; NC, negative control, no DNA template.

Mentions: The immunoreactivity to Cre was strongly observed in the cytoplasm of gastric, bronchial and intestinal epithelial cells (Figure 2C). To identify the exact cell types in which Cre recombinase performs its excision function, we bred K19-Cre transgenic mouse with the reporter mouse Rosa26-LacZ to activate β-galactosidase (Figure 3A). As shown in Figure 3B, galactosidase was detectable in gastric, bronchial and intestinal epithelial cells using x-gal as subtract. Wild-type C57 mouse (WT) was employed as a negative control and the target knockout mice of B6.Cg-Dkk 3tm1, B6.Cg-Becn1tm1, pvillin-Cre/Rosa as positive control due to the existence of endogenous LaZ gene respectively.


Cytokeratin 19 promoter directs the expression of Cre recombinase in various epithelia of transgenic mice
Cre activity in various tissues of K19-Cre transgeneic miceAfter mated with Rosa26-lacZ mouse A., Cre activity in K19-Cre/Rosa transgenic mice was visualized in lung, stomach and intestine by lacZ staining B. Wild-type C57 mouse (WT) was employed as a negative control and the target knockout mice of B6.Cg-Dkk 3tm1, B6.Cg-Becn1tm1 pvillin-Cre/Rosa as a positive control due to the existence of endogenous LaZ gene. Note: PC, positive control, tail DNA of pvillin- Cre mouse or Rosa26-lacZ; NC, negative control, no DNA template.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5392329&req=5

Figure 3: Cre activity in various tissues of K19-Cre transgeneic miceAfter mated with Rosa26-lacZ mouse A., Cre activity in K19-Cre/Rosa transgenic mice was visualized in lung, stomach and intestine by lacZ staining B. Wild-type C57 mouse (WT) was employed as a negative control and the target knockout mice of B6.Cg-Dkk 3tm1, B6.Cg-Becn1tm1 pvillin-Cre/Rosa as a positive control due to the existence of endogenous LaZ gene. Note: PC, positive control, tail DNA of pvillin- Cre mouse or Rosa26-lacZ; NC, negative control, no DNA template.
Mentions: The immunoreactivity to Cre was strongly observed in the cytoplasm of gastric, bronchial and intestinal epithelial cells (Figure 2C). To identify the exact cell types in which Cre recombinase performs its excision function, we bred K19-Cre transgenic mouse with the reporter mouse Rosa26-LacZ to activate β-galactosidase (Figure 3A). As shown in Figure 3B, galactosidase was detectable in gastric, bronchial and intestinal epithelial cells using x-gal as subtract. Wild-type C57 mouse (WT) was employed as a negative control and the target knockout mice of B6.Cg-Dkk 3tm1, B6.Cg-Becn1tm1, pvillin-Cre/Rosa as positive control due to the existence of endogenous LaZ gene respectively.

View Article: PubMed Central - PubMed

ABSTRACT

Cytokeratin 19 (K19) is expressed in various differentiated cells, including gastric, intestinal and bronchial epithelial cells, and liver duct cells. Here, we generated a transgenic mouse line, K19-Cre, in which the expression of Cre recombinase was controlled by the promoter of K19. To test the tissue distribution and excision activity of Cre recombinase, K19-Cre transgenic mice were bred with Rosa26 reporter strain and a mouse strain that carries PTEN conditional alleles (PTENLoxp/Loxp). At mRNA level, Cre was strongly expressed in the stomach, lung and intestine, while in stomach, lung, and liver at protein level. The immunoreactivity to Cre was strongly observed the cytoplasm of gastric, bronchial and intestinal epithelial cells. Cre activity was detectable in gastric, bronchial and intestinal epithelial cells, according to LacZ staining. In K19-Cre/PTEN Loxp/Loxp mice, PTEN was abrogated in stomach, intestine, lung, liver and breast, the former two of which were verified by in situ PCR. There appeared breast cancer with PTEN loss. These data suggest that K19 promoter may be a useful tool to study the pathophysiological functions of cytokeratin 19-positive cells, especially gastrointestinal epithelial cells. Cell specificity of neoplasia is not completely attributable to the cell-specific expression of oncogenes and cell-specific loss of tumor suppressor genes.

No MeSH data available.


Related in: MedlinePlus