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Krt5 creert2

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Krt5-CreERT2 is a transgenic mouse model that expresses a tamoxifen-inducible Cre recombinase under the control of the keratin 5 (Krt5) promoter. This model allows for the conditional expression or deletion of target genes in Krt5-expressing cells in a time-controlled manner.

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Lab products found in correlation

Krt5 creert2 knock in mice, by Jackson ImmunoResearch (1 mentions) R26r eyfp, by Jackson ImmunoResearch (1 mentions) Lsl krasg12d, by Jackson ImmunoResearch (1 mentions) Lgr5 egfp ires creert2, by Jackson ImmunoResearch (1 mentions) Krasg12d, by Addgene (1 mentions) Pik3cah1047r, by Addgene (1 mentions) Villin creert2, by Jackson ImmunoResearch (1 mentions) Sftpccreert2, by Jackson ImmunoResearch (1 mentions) C57bl 6j mice, by Japan SLC (1 mentions) Tgfbr2flox flox mice, by Jackson ImmunoResearch (1 mentions) Tamoxifen, by Merck Group (1 mentions) 4 hydroxytamoxifen 4 oht, by Merck Group (1 mentions)

6 protocols using krt5 creert2

1

Krt5-CreERT2:R26R-EYFP Lineage Tracing

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All the experiments were approved by the ethical committee from the university and conform with regulatory standards (LA1230406, project 666N). Mice were bred and maintained under pathogen-free conditions in a certified animal facility in accordance with the European guidelines. Adult (8-week-old) wild-type CD1 mice or Krt8-YFP knock-in mice (18 (link)) were used for sequencing and histology. Krt5-CreERT2 knock-in mice (the Jackson Laboratory, stock no. 029155; RRID:IMSR_JAX:029155) were crossed with R26R-EYFP (the Jackson Laboratory, stock no. 006148; RRID:IMSR_JAX:006148) to generate Krt5-CreERT2:R26R-EYFP (K5:RYFP) mice. For lineage tracing experiments, 10 or 0.25 mg of TAM resuspended in sunflower oil was injected intraperitoneally to 8-week-old K5:RYFP mice, and the expression of the YFP was analyzed in esophagi up to 4 weeks after TAM administration. In all the experiments, littermates of the same sex were randomly assigned to experimental groups.
Vercauteren Drubbel A, & Beck B. (2023). Single-cell transcriptomics uncovers the differentiation of a subset of murine esophageal progenitors into taste buds in vivo. Science Advances, 9(10), eadd9135.
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2

Generating Inducible Transgenic Mouse Models

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All inducible Cre lines (R26R-CreERT2: JAX006965, Villin-CreERT2: JAX020282, Lgr5-EGFP-IRES-CreERT2: JAX008875, Sftpc-CreERT2: JAX028054, Krt5-CreERT2: JAX02915), the R26R-Confetti line (JAX017492), LSL-KrasG12D (JAX008179) line, Apcfl/fl line (JAX009045) and ApcMin line (JAX002020) were obtained from The Jackson Laboratory. Pik3caLat-H1047R line was donated by W.A Phillips41 (link). Red2Onco targeting vectors were generated by gene synthesis. Oncogene sequences were obtained from Addgene (Notch1ICD: Addgene plasmid #1507942 (link), KrasG12D: Addgene plasmid #1154943 (link), PIK3CAH1047R: Addgene plasmid #1252444 (link)). Mice were created by inserting the Red2Onco cassette into the tdimer2 locus in ES cells obtained from R26R-Confetti mice using CRISPR-Cas9 nickase-mediated homologous recombination. Insertion of the oncogenic sequence was confirmed by long-range PCR. Specific genotyping primers were designed outside of the homology arms and were used in combination with primers within the knock-in cassette.
Yum M.K., Han S., Fink J., Wu S.H., Dabrowska C., Trendafilova T., Mustata R., Chatzeli L., Azzarelli R., Pshenichnaya I., Lee E., England F., Kim J.K., Stange D.E., Philpott A., Lee J.H., Koo B.K, & Simons B.D. (2021). Tracing oncogene-driven remodelling of the intestinal stem cell niche. Nature, 594(7863), 442-447.
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3

Genetic Manipulation of Tfap2c in Mice

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All animal studies conformed to guidelines set by the Office of the Institutional Animal Care and Use Committee (IACUC) and performed with IACUC approval. FVB/NJ and KRT5-Cre-ERT2 (FVB.Cg-Tg(Krt5-cre/ERT2)2Ipc/JeldJ) mice were purchased from Jackson Laboratory (stock no. 001800 and 018394). The Tfap2cfl/fl mouse strain was described previously (Auman et al., 2002 (link)) and was backcrossed >15 generations on FVB/NJ background to limit genetic variability. Knockout mice were generated by crossing Tfap2cfl/fl with Krt5-Cre-ERT2 mice, and the offspring were back crossed with Tfap2cfl/fl to generate FVB/Tfap2cfl/fl/Krt5-Cre-ERT2. Pulsing with 4OHT was performed starting at age 4 weeks at 30 mg/kg/day intraperitoneal injection for 5 days. All experimental analyses were performed in female mice. Genotype analysis was conducted with qPCR analysis of tail DNA by Transnetyx (Cordova, TN).
Gu V.W., Cho E., Thompson D.T., Cassady V.C., Borcherding N., Koch K.E., Wu V.T., Lorenzen A.W., van der Heide D.M., White J.R., Kulak M.V., Williams T., Zhang W, & Weigel R.J. (2020). AP-2γ Is Required for Maintenance of Multipotent Mammary Stem Cells. Stem Cell Reports, 16(1), 106-119.
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4

Genetic Lineage Tracing in Mouse Lung

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Krt5-CreERT2 (The Jackson Laboratory), Rosa26R-CAG-Confetti (Rosa-Confetti; supplied by Hans Clevers), and Foxj1-GFP (Ostrowski et al., 2003 (link)) mice were maintained on a C57BL/6 background. C57BL/6J mice were purchased from Japan SLC (Shizuoka, Japan). Male mice at 8–12 weeks old were used for SO2 injury. All experiments were performed in accordance with Yokohama City University Institutional Guidelines for Laboratory Animal Usage and IACUC-approved protocols at the Duke University School of Medicine.
Tadokoro T., Tanaka K., Osakabe S., Kato M., Kobayashi H., Hogan B.L, & Taniguchi H. (2021). Dorso-ventral heterogeneity in tracheal basal stem cells. Biology Open, 10(9), bio058676.
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5

Bladder Cancer Mouse Model Development

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KRT5-CreERT2 and Tgfbr2flox/flox mice were obtained from Jackson Laboratory, Balb/c male mice were provided by Shanghai Laboratory Animals Center (SLAC). BBN (Santa Cruz) was dissolved in drinking water at a concentration of 0.05% (w/v) and provided to transgenic or wild type male mice for 30 weeks as previous described38 (link). Tamoxifen (T5648) were purchased from Sigma, BBN(sc-486264) and LY 364947 (sc-203122) were obtained from Santa Cruz. All animal procedures were performed under a protocol approved by the Laboratory Animal Center of Anhui Medical University and in accordance with National Institutes of Health guide for the care and use of Laboratory animals (NIH Publications No. 8023, revised 1978).
Liang Y., Zhu F., Zhang H., Chen D., Zhang X., Gao Q, & Li Y. (2016). Conditional ablation of TGF-β signaling inhibits tumor progression and invasion in an induced mouse bladder cancer model. Scientific Reports, 6, 29479.
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6

Conditional Knockout of Ube2n in Skin

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Animal studies were conducted in accordance with protocols approved by Duke Animal Care and Use Committee. All mice were maintained under a temperature controlled and specific pathogen-free environment. Krt5CreERT2 (strain #: 029155; (75 (link))) was obtained from Jackson laboratories (Bar Harbor, ME). Rosa26CreER.Ube2nfl/fl mice were kindly gifted by Dr. Shao-Cong Sun (University of Texas MD Anderson, Houston, TX) under permission of Dr. Shizuo Akira (Osaka University, Osaka, Japan) (17 (link)). The Krt5CreER.Ube2nfl/fl was generated via crossbreeding and genotyped via PCR with primers listed in Table S2. Both male and female mice were used in this study. To induce local deletion of Ube2n in the skin, 1–6 months-old adult mice were shaved on the dorsal skin and treated topically with 40 μl of 1–5mg/ml 4-hydroxytamoxifen (4-OHT; Sigma, St. Louis, MO) in ethanol every other day for 4 times. For the time-course experiment, 20 μl of 1 mg/ml of 4-OHT was applied daily for 4 days. The vehicle control mice were treated with ethanol. For the WT control, littermates Ube2nfl/fl mice were treated with equal amounts of 4-OHT.
Lee M.J., Hammouda M.B., Miao W., Okafor A., Jin Y., Sun H., Jain V., Markovtsov V., Diao Y., Gregory S.G, & Zhang J.Y. (2023). UBE2N is essential for maintenance of skin homeostasis and suppression of inflammation. bioRxiv.
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