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The generation and characterization of novel Col1a1FRT-Cre-ER-T2-FRT and Col1a1FRT-STOP-FRT-Cre-ER-T2 mice for sequential mutagenesis.

Zhang M, Kirsch DG - Dis Model Mech (2015)

Bottom Line: This application of dual recombinase technology can be used to dissect the role of stromal cells in tumor development and cancer therapy.To potentially utilize the large number of Cre-loxP-regulated transgenic alleles that have already been targeted into the Rosa26 locus, such as different reporters and mutant genes, we targeted the two novel Cre-ER(T2) alleles into the endogenous Col1a1 locus for ubiquitous expression.These two new novel mouse strains will be complementary to each other and will enable the exploration of complex biological questions in development, normal tissue homeostasis and cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Cancer Biology, Duke University, Durham, NC 27708, USA.

No MeSH data available.


Related in: MedlinePlus

Characterization of tamoxifen-independent Cre-ERT2 activity in Col1a1FRT-Cre-ER-T2-FRT mice. (A) Immunofluorescence of tissues from 4-month-old Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice (n=2) without tamoxifen treatment show time- and tissue-dependent leakiness of Cre-ERT2, with more widespread eGFP expression in the skeletal muscle (Aiii), pancreas (Avii) and liver (Axv). (B) Immunofluorescence of tissues from 6-month-old Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice (n=3) without tamoxifen treatment show increased time- and tissue-dependent leakiness of Cre-ERT2, with more widespread eGFP expression in the skeletal muscle (Biii), pancreas (Bvii) and liver (Bxv). The skeletal muscle and the pancreas show the most apparent increase in eGFP expression from 4 to 6 months. Scale bars: 50 µm.
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DMM021204F5: Characterization of tamoxifen-independent Cre-ERT2 activity in Col1a1FRT-Cre-ER-T2-FRT mice. (A) Immunofluorescence of tissues from 4-month-old Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice (n=2) without tamoxifen treatment show time- and tissue-dependent leakiness of Cre-ERT2, with more widespread eGFP expression in the skeletal muscle (Aiii), pancreas (Avii) and liver (Axv). (B) Immunofluorescence of tissues from 6-month-old Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice (n=3) without tamoxifen treatment show increased time- and tissue-dependent leakiness of Cre-ERT2, with more widespread eGFP expression in the skeletal muscle (Biii), pancreas (Bvii) and liver (Bxv). The skeletal muscle and the pancreas show the most apparent increase in eGFP expression from 4 to 6 months. Scale bars: 50 µm.

Mentions: To determine the activity of Cre-ERT2 in Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice in the absence of tamoxifen, tissues from 4-month-old (n=2) and 6-month-old (n=3) mice without tamoxifen treatment were examined (Fig. 5). The tissues with the most widespread eGFP expression without tamoxifen treatment in the 6-month-old mice include the pancreas (Fig. 5Bvii), the liver (Fig. 5Bxv) and the skeletal muscle (Fig. 5Biii). There was a time-dependent tissue-specific increase in eGFP expression when comparing the 4-month-old and 6-month-old mice (Fig. 5A,B). For example, the skeletal muscle fibers in the 4-month-old mice (Fig. 5Aiii) had less eGFP expression than those in the 6-month-old mice (Fig. 5Biii). Additionally, the pancreas of the 4-month-old mice (Fig. 5Avii) had less eGFP expression than the pancreas of the 6-month-old mice (Fig. 5Bvii). The liver of both the 4-month-old (Fig. 5Axv) and the 6-month-old (Fig. 5Bxv) mice had similar eGFP expression. In contrast, the brain had no eGFP expression with the exception of a subset of the vasculature in both the 4-month-old mice (Fig. 5Axxxi) and the 6-month-old mice (Fig. 5Bxxxi). Taken together, these results demonstrate that there is time-dependent and tissue-specific Cre-ERT2 activity in Col1a1FRT-Cre-ER-T2-FRT mice in the absence of tamoxifen, with the brain tissue being the least affected organ, and the pancreas, liver and skeletal muscle fibers being the most affected organs. In addition, tamoxifen potently induces Cre-ERT2-mediated recombination of unrecombined loxP sites.Fig. 5.


The generation and characterization of novel Col1a1FRT-Cre-ER-T2-FRT and Col1a1FRT-STOP-FRT-Cre-ER-T2 mice for sequential mutagenesis.

Zhang M, Kirsch DG - Dis Model Mech (2015)

Characterization of tamoxifen-independent Cre-ERT2 activity in Col1a1FRT-Cre-ER-T2-FRT mice. (A) Immunofluorescence of tissues from 4-month-old Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice (n=2) without tamoxifen treatment show time- and tissue-dependent leakiness of Cre-ERT2, with more widespread eGFP expression in the skeletal muscle (Aiii), pancreas (Avii) and liver (Axv). (B) Immunofluorescence of tissues from 6-month-old Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice (n=3) without tamoxifen treatment show increased time- and tissue-dependent leakiness of Cre-ERT2, with more widespread eGFP expression in the skeletal muscle (Biii), pancreas (Bvii) and liver (Bxv). The skeletal muscle and the pancreas show the most apparent increase in eGFP expression from 4 to 6 months. Scale bars: 50 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4582108&req=5

DMM021204F5: Characterization of tamoxifen-independent Cre-ERT2 activity in Col1a1FRT-Cre-ER-T2-FRT mice. (A) Immunofluorescence of tissues from 4-month-old Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice (n=2) without tamoxifen treatment show time- and tissue-dependent leakiness of Cre-ERT2, with more widespread eGFP expression in the skeletal muscle (Aiii), pancreas (Avii) and liver (Axv). (B) Immunofluorescence of tissues from 6-month-old Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice (n=3) without tamoxifen treatment show increased time- and tissue-dependent leakiness of Cre-ERT2, with more widespread eGFP expression in the skeletal muscle (Biii), pancreas (Bvii) and liver (Bxv). The skeletal muscle and the pancreas show the most apparent increase in eGFP expression from 4 to 6 months. Scale bars: 50 µm.
Mentions: To determine the activity of Cre-ERT2 in Col1a1FRT-Cre-ER-T2-FRT; Rosa26mTmG/+ mice in the absence of tamoxifen, tissues from 4-month-old (n=2) and 6-month-old (n=3) mice without tamoxifen treatment were examined (Fig. 5). The tissues with the most widespread eGFP expression without tamoxifen treatment in the 6-month-old mice include the pancreas (Fig. 5Bvii), the liver (Fig. 5Bxv) and the skeletal muscle (Fig. 5Biii). There was a time-dependent tissue-specific increase in eGFP expression when comparing the 4-month-old and 6-month-old mice (Fig. 5A,B). For example, the skeletal muscle fibers in the 4-month-old mice (Fig. 5Aiii) had less eGFP expression than those in the 6-month-old mice (Fig. 5Biii). Additionally, the pancreas of the 4-month-old mice (Fig. 5Avii) had less eGFP expression than the pancreas of the 6-month-old mice (Fig. 5Bvii). The liver of both the 4-month-old (Fig. 5Axv) and the 6-month-old (Fig. 5Bxv) mice had similar eGFP expression. In contrast, the brain had no eGFP expression with the exception of a subset of the vasculature in both the 4-month-old mice (Fig. 5Axxxi) and the 6-month-old mice (Fig. 5Bxxxi). Taken together, these results demonstrate that there is time-dependent and tissue-specific Cre-ERT2 activity in Col1a1FRT-Cre-ER-T2-FRT mice in the absence of tamoxifen, with the brain tissue being the least affected organ, and the pancreas, liver and skeletal muscle fibers being the most affected organs. In addition, tamoxifen potently induces Cre-ERT2-mediated recombination of unrecombined loxP sites.Fig. 5.

Bottom Line: This application of dual recombinase technology can be used to dissect the role of stromal cells in tumor development and cancer therapy.To potentially utilize the large number of Cre-loxP-regulated transgenic alleles that have already been targeted into the Rosa26 locus, such as different reporters and mutant genes, we targeted the two novel Cre-ER(T2) alleles into the endogenous Col1a1 locus for ubiquitous expression.These two new novel mouse strains will be complementary to each other and will enable the exploration of complex biological questions in development, normal tissue homeostasis and cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Cancer Biology, Duke University, Durham, NC 27708, USA.

No MeSH data available.


Related in: MedlinePlus