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Mmtv cre

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MMTV-Cre is a genetic construct that expresses the Cre recombinase enzyme under the control of the mouse mammary tumor virus (MMTV) promoter. The Cre recombinase enzyme is a site-specific DNA recombinase that can be used to modify or excise DNA sequences in transgenic mice or cell lines.

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Lab products found in correlation

Tcfap2cl, by Jackson ImmunoResearch (2 mentions) Eif2ak3tm1.2drc, by Jackson ImmunoResearch (2 mentions) Tg mmtv cre 4mam, by Jackson ImmunoResearch (2 mentions) Her2 neu transgenic mice, by Jackson ImmunoResearch (2 mentions) Mx cre mice, by Jackson ImmunoResearch (1 mentions) Krt14 cre, by Jackson ImmunoResearch (1 mentions) Tg mmtv cre 4mam j, by Jackson ImmunoResearch (1 mentions) Polyinosinic polycytidylic acid pipc, by Merck Group (1 mentions)

12 protocols using mmtv cre

1

Conditional Foxa1 Deletion in Mouse Mammary Gland

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All animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) at the Mayo Clinic. The experiment was carried out under controlled conditions with a 12-h light/dark cycle. Cages with filters were used along with sterile bedding, ad libitum diet, and water. Animals were maintained on a normal chow. The derivation of Foxa1loxP/loxP mice (from Dr. Klaus H. Kaestner, University of Pennsylvania) has been reported previously (Gao et al., 2008a (link)). Foxa1loxP/loxP mice were mated with MMTV-Cre (from Jackson Laboratory), MMTV-rtTA;Tet-On-Cre (from Dr. Lewis A. Chodosh, University of Pennsylvania) (Gunther et al., 2002 (link)), or Krt14-Cre (from Jackson Laboratory) mice to obtain Foxa1loxP/loxP;MMTV-Cre, Foxa1loxP/loxP;MMTV-rtTA;Tet-On-Cre, or Foxa1loxP/loxP;Krt14-Cre mice. Foxa1loxP/loxP mice were used as control wild-type (WT) mice. Genotypes of Foxa1loxP/loxP and Cre were determined by PCR. 2 μg/ml doxycycline in drinking water was given to Foxa1loxP/loxP;MMTV-rtTA;Tet-On-Cre mice from age of 6 weeks old to 12 weeks old to induce Cre expression. Mammary glands of all female mice (3 to 10 mice per group) were collected at 12 weeks old for the following analysis.
Liu Y., Zhao Y., Skerry B., Wang X., Colin-Cassin C., Radisky D.C., Kaestner K.H, & Li Z. (2016). Foxa1 is Essential for Mammary Duct Formation. Genesis (New York, N.Y. : 2000), 54(5), 277-285.
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2

Generating Conditional Knockout Mouse Models

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All animal work was performed according to institutional regulations for care and use of laboratory animals and approved by the Lower Saxony State Office for Consumer Protection and Food Safety (LAVES; registration numbers: G15/2039, G15/1754 and G11/540). Usp22flox mice were generated as described previously6 by removing lacZ and neomycin resistance loci from the Usp22tm1a(KOMP)Wtsi C57BL6 mouse line generated from embryonic stem cells obtained from the University of California-Davis Knockout Mouse Project Repository20 (link) by FLP-mediated excision21 (link). Usp22flox mice were crossed with MMTV-Cre (a kind gift from L. Henninghausen, National Institutes of Health, USA) and Tg(MMTV-ErbB2)NK1Mul/J (The Jackson Laboratory) animals (FVB/N background) to achieve a mammary-specific Usp22 knockout and to promote tumorigenesis, respectively. Moreover, Usp22flox mice were crossed with Villin-CreERT2 and Apc1638N mouse lines (C57BL/6N background) to achieve an intestinal knockout and to investigate its role in tumorigenesis6 . The Apc1638N/+ mouse line was a kind gift from F. Bosman (Erasmus University Medical Center Rotterdam, The Netherlands). Multiple replicates (n = 3–6) were utilized in order to ensure reproducibility of findings.
Kosinsky R.L., Helms M., Zerche M., Wohn L., Dyas A., Prokakis E., Kazerouni Z.B., Bedi U., Wegwitz F, & Johnsen S.A. (2019). USP22-dependent HSP90AB1 expression promotes resistance to HSP90 inhibition in mammary and colorectal cancer. Cell Death & Disease, 10(12), 911.
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3

Tcfap2c Regulates Mammary Tumor Development

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All animal protocols adhered to guidelines by the University of Iowa Institutional Animal Care and Use Committee. Mice harboring loxP-flanked Tcfap2c alleles were obtained from Dr Trevor Williams and maintained as described.14 (link) Mice harboring the MMTV-Neu allele were obtained from Dr Bill Muller. Mice harboring the MMTV-Cre allele were purchased from Jackson Laboratories (Bar Harbor, ME, USA). F4 progeny carrying Tcfap2c allele carrying or lacking MMTV-Cre were crossed with female mice harboring MMTV-Neu. These progeny were used for analysis. Genotyping was carried out with tail snips and analyzed by Transnetyx (Cordova, TN, USA). Tumors (n = 3 tumors) were harvested from mice, formalin fixed and paraffin embedded. Hematoxylin and eosin and IHC analyses were completed with the aid from the University of Iowa Pathology Core Lab with the aid from a veterinary pathologist. Categorical data were analyzed by Fisher's exact test. IHC was analyzed by t-test. Survival curves were analyzed by long-rank test.
Park J., Wu T., Cyr A., Woodfield G., De Andrade J., Spanheimer P., Li T., Sugg S., Lal G., Domann F., Zhang W, & Weigel R. (2015). The role of Tcfap2c in tumorigenesis and cancer growth in an activated Neu model of mammary carcinogenesis. Oncogene, 34(50), 6105-6114.
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4

Genetic Analysis of Mammary Gland Development

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Mice harboring the exon 6 LoxP-flanked Tcfap2c (Tcfap2cL/L) allele were obtained from Dr. Trevor Williams, and mice harboring the MMTV-Cre allele were purchased from Jackson Laboratories (003553, Bar Harbor, ME, USA) 35 . All animals used for studies were from F4 progeny breeds to limit genetic variability. Genotyping was performed by Transnetyx, Inc (Cordova, TN, USA) on tissue harvested from tail-snip biopsy. All animals were cared for in accordance with guidelines established by the University of Iowa Institutional Animal Care and Use Committee. Whole mounts were accomplished as previously described according to Plante et al, using mammary gland 4 35 . Immunohistochemical analysis was accomplished on FFPE samples from mammary gland 3. Data from whole mount testing was assessed using Student’s 2-tailed T-test.
Cyr A.R., Kulak M.V., Park J.M., Bogachek M.V., Spanheimer P.M., Woodfield G.W., White-Baer L.S., O’Malley Y.Q., Sugg S.L., Olivier A.K., Zhang W., Domann F.E, & Weigel R.J. (2014). TFAP2C Governs the Luminal Epithelial Phenotype in Mammary Development and Carcinogenesis. Oncogene, 34(4), 436-444.
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5

Generation of Fatty Acid-Deficient Mammary Milk

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The Protein Kinase R (PKR)-like endoplasmic reticulum kinase (PERK) or
Eukaryotic translation initiation factor 2 alpha-subunit kinase 3 (Eif2ak3)
plays a significant role in the unfolded protein response55 (link),56 (link). PERK is required for proper development and maturation of
mammary epithelial cells. Deletion of PERK in mammary glands inhibits key
lipogenic enzymes involved in lipid production and storage (Fatty acid synthase,
ATP citrate lyase and stearyl-CoA desaturase-1) decreasing the lipid content in
milk45 (link). To generate
mice which produce milk deficient in fatty acids, PERKloxP mice with
loxP sites which flank exons 3–557 (link) were mated with Mouse mammary tumor virus (MMTV)-Cre
transgenic mice46 (link). MMTV-cre
transgenic mouse lines express Cre recombinase predominantly in secretory cell
types in the mammary epithelium46 (link). Female mice homozygous for PERKloxP and
MMTV-cre produce mammary glands in which the milk is deficient in fatty
acids45 (link). Here, only
male mice were used in our studies to avoid variations between groups. The
PERKloxP and MMTV-cre mice were obtained from Jackson Laboratory
(Eif2ak3tm1.2Drc stock number 023066); and Tg(MMTV-cre)4Mam -
stock number 003553), respectively.
Cardoso A.C., Lam N.T., Savla J.J., Nakada Y., Pereira A.H., Elnwasany A., Menendez-Montes I., Ensley E.L., Petric U.B., Sharma G., Sherry A.D., Malloy C.R., Khemtong C., Kinter M.T., Tan W.L., Anene-Nzelu C.G., Foo R.S., Nguyen N.U., Li S., Ahmed M.S., Elhelaly W.M., Abdisalaam S., Asaithamby A., Xing C., Kanchwala M., Vale G., Eckert K.M., Mitsche M.A., McDonald J.G., Hill J.A., Huang L., Shaul P.W., Szweda L.I, & Sadek H.A. (2020). Mitochondrial Substrate Utilization Regulates Cardiomyocyte Cell Cycle Progression. Nature metabolism, 2(2), 167-178.
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6

Generation and Characterization of RANK Knockout Mice

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Example 1

Mice.

Rankfloxed mice have been recently generated1. Briefly, to generate mice carrying a null allele of Rank (rankΔ allele), rankfloxed mice were crossed to β-actin-Cre ubiquitous deleter mice. Mice carrying the rankfloxed or rankΔ alleles as well as the MMTV-Cre mice were backcrossed seven times onto a BALBc background before generating the MMTV-Cre rankΔ/floxed mice. MMTV-NeuT mice were kindly provided by Guido Forni, Milan. MMTV-Cre (stock #003553) and Mx-Cre mice (stock #003556) were obtained from the Jackson Laboratory. K5-Cre, IKKαfloxed and NFATc1floxed mice have been previously described2-4. Mouse geno-types were determined by PCR and Southern blot analysis. In all experiments, only littermate mice from the same breedings were used. All mice were bred and maintained according to institutional guidelines.

RANK Deletion in Tumors and Cre Effects.

Southern blotting of the tumors that developed in RANKΔmam females showed deletion of RANK, albeit some residual wild type band was observed (FIG. 10c) that may be explained by the presence of other cell types and/or escaper cells. Differences in tumor onset in Cre-negative control females and littermates expressing the MMTV-Cre transgene were not observed indicating that Cre expression per se does not alter tumor incidence in the MPA/DMBA mammary tumor model (FIG. 10d).

US10143747B2. Breast cancer therapeutics (2018-12-04). None [AT], None [AT]. Inventors: Josef Penninger [AT], Daniel Schramek [AT].
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7

Generation of Fatty Acid-Deficient Mammary Milk

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The Protein Kinase R (PKR)-like endoplasmic reticulum kinase (PERK) or
Eukaryotic translation initiation factor 2 alpha-subunit kinase 3 (Eif2ak3)
plays a significant role in the unfolded protein response55 (link),56 (link). PERK is required for proper development and maturation of
mammary epithelial cells. Deletion of PERK in mammary glands inhibits key
lipogenic enzymes involved in lipid production and storage (Fatty acid synthase,
ATP citrate lyase and stearyl-CoA desaturase-1) decreasing the lipid content in
milk45 (link). To generate
mice which produce milk deficient in fatty acids, PERKloxP mice with
loxP sites which flank exons 3–557 (link) were mated with Mouse mammary tumor virus (MMTV)-Cre
transgenic mice46 (link). MMTV-cre
transgenic mouse lines express Cre recombinase predominantly in secretory cell
types in the mammary epithelium46 (link). Female mice homozygous for PERKloxP and
MMTV-cre produce mammary glands in which the milk is deficient in fatty
acids45 (link). Here, only
male mice were used in our studies to avoid variations between groups. The
PERKloxP and MMTV-cre mice were obtained from Jackson Laboratory
(Eif2ak3tm1.2Drc stock number 023066); and Tg(MMTV-cre)4Mam -
stock number 003553), respectively.
Cardoso A.C., Lam N.T., Savla J.J., Nakada Y., Pereira A.H., Elnwasany A., Menendez-Montes I., Ensley E.L., Petric U.B., Sharma G., Sherry A.D., Malloy C.R., Khemtong C., Kinter M.T., Tan W.L., Anene-Nzelu C.G., Foo R.S., Nguyen N.U., Li S., Ahmed M.S., Elhelaly W.M., Abdisalaam S., Asaithamby A., Xing C., Kanchwala M., Vale G., Eckert K.M., Mitsche M.A., McDonald J.G., Hill J.A., Huang L., Shaul P.W., Szweda L.I, & Sadek H.A. (2020). Mitochondrial Substrate Utilization Regulates Cardiomyocyte Cell Cycle Progression. Nature metabolism, 2(2), 167-178.
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8

Genetic Analysis of Mammary Gland Development

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Mice harboring the exon 6 LoxP-flanked Tcfap2c (Tcfap2cL/L) allele were obtained from Dr. Trevor Williams, and mice harboring the MMTV-Cre allele were purchased from Jackson Laboratories (003553, Bar Harbor, ME, USA) 35 . All animals used for studies were from F4 progeny breeds to limit genetic variability. Genotyping was performed by Transnetyx, Inc (Cordova, TN, USA) on tissue harvested from tail-snip biopsy. All animals were cared for in accordance with guidelines established by the University of Iowa Institutional Animal Care and Use Committee. Whole mounts were accomplished as previously described according to Plante et al, using mammary gland 4 35 . Immunohistochemical analysis was accomplished on FFPE samples from mammary gland 3. Data from whole mount testing was assessed using Student’s 2-tailed T-test.
Cyr A.R., Kulak M.V., Park J.M., Bogachek M.V., Spanheimer P.M., Woodfield G.W., White-Baer L.S., O’Malley Y.Q., Sugg S.L., Olivier A.K., Zhang W., Domann F.E, & Weigel R.J. (2014). TFAP2C Governs the Luminal Epithelial Phenotype in Mammary Development and Carcinogenesis. Oncogene, 34(4), 436-444.
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9

Crossbreeding Protocol for Mouse Models

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The SHP2-floxed mice (25 (link)) were obtained from Dr. Benjamin Neel, the HER2/Neu transgenic mice (11 (link)) were purchased from the Jackson Laboratory, and the MMTV-Cre (26 (link)) were obtained from Dr. Timothy Lane. A standard crossbreeding protocol as outlined in Figure 1a was used in the study. Genomic DNA isolated from tail snips was used for PCR genotyping of mice. The forward and the reverse primers in a respective order include 5’-TAGCTGCTTTAACCCTCTGTGT-3’ and 5’-CATCAGADCAGGCCATATTCC-3’ for SHP2, 5’-TTTCCTGCAGCAGCCTACGC-3’ and 5’CGGAACCCACATCAGGCC-3’ for HER2, and 5’-ATCCGAAAAGAAAACGTTGA-3’ and 5’-ATCCAGCTTACGGATATAGT-3’ for Cre. The internal control primers for the HER2 transgene were the forward 5’-CAAATGTTGCTTGTCTGGTG-3’ and the reverse 5’-GTCAGTCGAGTGCACAGTTT-3’. All genetic studies were conducted by strictly following the approved protocol by the West Virginia University Institutional Committee for animal Use and Care (WVU ICUC).
Zhao H., Martin E., Matalkah F., Shah N., Ivanov A., Ruppert J.M., Lockman P.R, & Agazie Y.M. (2018). Conditional knockout of SHP2 in ErbB2 transgenic mice or inhibition in HER2-amplified breast cancer cell lines blocks oncogene expression and tumorigenesis. Oncogene, 38(13), 2275-2290.
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10

Conditional Loxl2 Modulation in MMTV-PyMT Mice

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MMTV-PyMT (PyMT) mice (FBV/n background) were purchased from The Jackson laboratory and MMTV-Cre (26 (link)) were obtained from the CNIO (Centro Nacional de Investigaciones Oncológicas) Transgenic Unit. Conditional Loxl2 deletion (L2fl/fl) or overexpression (R26STOPL2) mouse lines were recently described (22 (link)); presenting a mixed genetic background (50% FVB and 50% mix from C57BL/6, CD1 and 129v strains). PyMT mice harboring conditional Loxl2 deletion (PyMT;L2Δ/-) or overexpression (PyMT;R26L2) in the mammary gland were generated by intercrossing L2fl/fl or R26STOPL2 females with their corresponding MMTV-PyMT+/T;MMTV-Cre+/T;Loxl2fl/- and MMTV-PyMT+/T;MMTV-Cre+/T;R26STOPL2 males. PyMT+/T;L2fl/fl and PyMT+/T;R26STOPL2 mice lacking Cre expression (Cre+/+) were used as littermate controls. All mouse studies were conducted in accordance with protocols approved by the Use Committee for Animal Care from the Universidad Autónoma de Madrid (UAM) (Ref# CEI-25-587) and the Comunidad de Madrid (PROEX 182/14).
Salvador F., Martin A., López-Menéndez C., Moreno-Bueno G., Santos V., Vázquez-Naharro A., Santamaria P.G., Morales S., Dubus P.R., Muinelo-Romay L., López-López R., Tung J.C., Weaver V.M., Portillo F, & Cano A. (2017). Lysyl oxidase-like protein LOXL2 promotes lung metastasis of breast cancer. Cancer research, 77(21), 5846-5859.
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