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Rosa26 creert2 mice

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The Rosa26-CreERT2 mice are a transgenic mouse model that expresses a tamoxifen-inducible Cre recombinase under the control of the ubiquitously expressed Rosa26 promoter. This allows for temporal and spatial control of Cre-mediated gene recombination.

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Lab products found in correlation

Mx1 cre, by Jackson ImmunoResearch (2 mentions) Gli1 cre ert2 mice, by Jackson ImmunoResearch (1 mentions) C57bl 6 wild type mice, by Janvier Labs (1 mentions) C57bl 6 mice, by BioXCell (1 mentions) C57bl 6 mice, by Jackson ImmunoResearch (1 mentions) Corn oil, by Merck Group (1 mentions) Tamoxifen corn oil solution, by Merck Group (1 mentions) Rag2 il2rg mice, by Taconic Biosciences (1 mentions) B6 sjl, by Taconic Biosciences (1 mentions) Rosa26 creert2, by Jackson ImmunoResearch (1 mentions) Tamoxifen, by Jackson ImmunoResearch (1 mentions) Tamoxifen, by Merck Group (1 mentions)

13 protocols using rosa26 creert2 mice

1

Generation and Characterization of Transgenic Mouse Models

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Generation of Kit-KO, Kit-EGFP, and Sox2loxP/loxP mice have been previously described [8 (link), 37 (link), 41 (link)]. Sox2loxP/loxP were intercrossed with Rosa26Cre-Ert2 mice (Jackson Laboratories, Ann Harbor, MI) to generate Sox2loxP/loxP, Rosa26Cre-Ert2 mice. Sox2loxP/loxP Rosa26Cre-ERT mice were crossed to p18 mice to generate Sox2loxP/loxP, Rosa26Cre-Ert2 Kit-EGFP (p18) mice. Kit-KO mice were intercrossed with p18 to generate Kit-KO, Kit-EGFP. For embryo staging, 0.5 dpc corresponded to the day of vaginal plug. Knockin mice carrying the KitD814Y mutation (see below) were generated by injecting mutant ESCs into C57/B6 carrier blastocysts and transplanted the same day into foster CD1 mothers. All experiments were performed in compliance with the Tor Vergata University Institutional Animal Care and National Institutes of Health Intramural Animal Care and Use program. All procedures adhered to the standards published in Guide for the Care and Use of Laboratory Animals. Experimental procedures involving mice were approved by the Italian Ministry of Health. The structure of the targeting construct used to generate KitD814Y knockin ESCs is depicted below.
Todaro F., Campolo F., Barrios F., Pellegrini M., Di Cesare S., Tessarollo L., Rossi P., Jannini E.A, & Dolci S. (2019). Regulation of Kit Expression in Early Mouse Embryos and ES Cells. Stem cells (Dayton, Ohio), 37(3), 332-344.
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2

Generation of H3.1-iCOUNT Transgenic Mice

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C57BL/6 mice expressing H3.1-iCOUNT were generated by PolyGene (Switzerland). C57BL/6-derived ESCs were transfected with the iCOUNT cassette targeting H3.1 as described above and selected using G418. Single clones were picked, expanded and correct insertion was verified by PCR. Positive cells were injected into blastocysts and 6 chimeras were born. Crossing three chimeras to gray C57BL/6 females yielded heterozygous H3.1-iCOUNT positive animals. To generate homozygous H3.1-iCOUNT mice, H3.1-iCOUNT ± mice were crossed with each other. Alternatively, H3.1-iCOUNT animals were crossed to Gli1:Cre-ERT2 mice (Jackson Lab No 007913: (Ahn and Joyner, 2004 (link)) and ROSA26:Cre-ERT2 mice (Jackson Lab No 008463; (Ventura et al., 2007 (link)). Animals were always genotyped to distinguish WT, heterozygous and homozygous H3.1-iCOUNT animals (see PCR above). H3.1 tagging caused mosaic expression of the RITE cassette, as there 9 different genes encoding for the histone-variants H3.1 and H3.2. within the HIST1 gene cluster on chromosome 13 (Wang et al., 1996 (link)). Indeed, we observed mosaic expression and mice that had the RITE cassette correctly inserted (verified via genomic PCR) but did not express the transgene.
Denoth-Lippuner A., Jaeger B.N., Liang T., Royall L.N., Chie S.E., Buthey K., Machado D., Korobeynyk V.I., Kruse M., Munz C.M., Gerbaulet A., Simons B.D, & Jessberger S. (2021). Visualization of individual cell division history in complex tissues using iCOUNT. Cell Stem Cell, 28(11), 2020-2034.e12.
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3

Tamoxifen-induced Klf4 Knockout Mice

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Klf4-floxed mice were obtained from The Mutant Mouse Regional Resource Center (MMRRC) at the University of Missouri, Columbia, Missouri, USA. Rosa26-CreERT2 mice were obtained from The Jackson Laboratory. Tamoxifen-inducible Klf4-knockout mice were generated by crossing Rosa26-CreERT2 mice and Klf4-floxed mice. Klf4 was knocked out by daily i.p. injection of tamoxifen at a dose of 100 mg/kg for 5 consecutive days. Wild-type C57BL/6 mice were obtained from Janvier Labs.
Satoh T.K., Mellett M., Meier-Schiesser B., Fenini G., Otsuka A., Beer H.D., Rordorf T., Maul J.T., Hafner J., Navarini A.A., Contassot E, & French L.E. (2020). IL-36γ drives skin toxicity induced by EGFR/MEK inhibition and commensal Cutibacterium acnes. The Journal of Clinical Investigation, 130(3), 1417-1430.
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4

LCMV Infection in C57BL/6 Mice

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C57BL/6 mice and Rosa26Cre-ERT2 mice were purchased from the Jackson Laboratory. The dLck-Cre TGFβRIIflox/flox (KO) mice, generously given by N. Zhang (University of Texas at San Antonio, San Antonio, TX), were crossed to LCMV DbGP33-specific TCR transgenic P14 mice in-house. To induce persistent LCMV infection, 6–8-wk-old female C57BL/6 mice were given anti-CD4–depleting antibody GK1.5 (Bio X Cell) i.p. twice before i.v. infection with 2 × 106 PFU LCMV clone 13 strain (Matloubian et al., 1994 (link)). All animal experiments were performed in accordance with the Emory University Institutional Animal Care and Use Committee.
Hu Y., Hudson W.H., Kissick H.T., Medina C.B., Baptista A.P., Ma C., Liao W., Germain R.N., Turley S.J., Zhang N, & Ahmed R. (2022). TGF-β regulates the stem-like state of PD-1+ TCF-1+ virus-specific CD8 T cells during chronic infection. The Journal of Experimental Medicine, 219(10), e20211574.
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5

Conditional Ptpn11 Knockout Mouse Model

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Ptpn11flox/flox mice41 (link),42 (link) were bred to Mx1-Cre or Rosa26-creERT2 mice (The Jackson Laboratory) in the C57BL/6 background. Genotyping was performed as described.31 (link)
Gu S., Sayad A., Chan G., Yang W., Lu Z., Virtanen C., Van Etten R.A, & Neel B.G. (2017). SHP2 Is Required for BCR-ABL1-Induced Hematologic Neoplasia. Leukemia, 32(1), 203-213.
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6

Conditional Ptpn11 Knockout Mouse Model

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Ptpn11flox/flox mice41 (link),42 (link) were bred to Mx1-Cre or Rosa26-creERT2 mice (The Jackson Laboratory) in the C57BL/6 background. Genotyping was performed as described.31 (link)
Gu S., Sayad A., Chan G., Yang W., Lu Z., Virtanen C., Van Etten R.A, & Neel B.G. (2017). SHP2 Is Required for BCR-ABL1-Induced Hematologic Neoplasia. Leukemia, 32(1), 203-213.
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7

Generation of H3.3A/B Knockout Mice

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The parental ESC line used to generate both H3.3A-and H3.3B-KO mice was derived from F1 hybrid embryos crossed with 129SV and C57BL/6J mice. The H3.3A-floxed ESC line was generated through knock-in of loxP sites to exon 3 of the h3f3a gene via a conventional recombinational approach. The H3.3B−/− ESC lines were generated through knock-in of a fluorescence (EYFP/mCherry) reporter and a stop codon to replace the three exons of the h3f3b gene via zinc finger nucleases. The mutant mice were produced through generation of chimaeras with the targeted ESCs, which were intercrossed with the germline transmission mice to obtain homozygous mutant mice. The CD45.1 NSG mice used for the BM transplantations were purchased from The Jackson Laboratory (strain 005557).
Rosa26creERT2+ mice were purchased from The Jackson Laboratory (https://www.jax.org/strain/008463). The Rosa26creERT2+; H3.3Afl/fl, H3.3B−/− breeder and offspring mice were maintained in a B6/129 background and backcrossed to C57BL/6J mice more than five times.
Guo P., Liu Y., Geng F., Daman A.W., Liu X., Zhong L., Ravishankar A., Lis R., Durán J.G., Itkin T., Tang F., Zhang T., Xiang J., Shido K., Ding B.S., Wen D., Josefowicz S.Z, & Rafii S. (2021). Histone variant H3.3 maintains adult haematopoietic stem cell homeostasis by enforcing chromatin adaptability. Nature cell biology, 24(1), 99-111.
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8

Tamoxifen-Induced CCN2 Knockout Mice

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Experiments were performed according to the European community guidelines for animal experiments and the ARRIVE guidelines and with consent of the Experimental Animal Ethics Committee of the University of Utrecht, The Netherlands [22 (link)]. Generation of tamoxifen-inducible CCN2 full-knockout mice is extensively described elsewhere [23 (link)]. In brief, CCN2Flox/Flox mice were crossbred with ROSA26CreERT2 mice (Gt(ROSA)26Sortm(cre/ERT2)Tyj/J, The Jackson Laboratory, Maine, USA), both on a C57Bl6/J background. Male 12–14 week old mice received four intraperitoneal injections on alternate days over a course of 7 days with either 100 μL corn oil (Sigma–Aldrich, St. Louis, MO, USA) or 100 μL of 10 mg/mL tamoxifen–corn oil solution (Sigma–Aldrich, St. Louis, MO, USA). After the last injection, a 14 day washout period was followed by the IRI operation. Sample sizes were based on published studies and pilot experiments. [24 (link)]
Valentijn F.A., Knoppert S.N., Pissas G., Rodrigues-Diez R.R., Marquez-Exposito L., Broekhuizen R., Mokry M., Kester L.A., Falke L.L., Goldschmeding R., Ruiz-Ortega M., Eleftheriadis T, & Nguyen T.Q. (2021). CCN2 Aggravates the Immediate Oxidative Stress–DNA Damage Response following Renal Ischemia–Reperfusion Injury. Antioxidants, 10(12), 2020.
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9

Genetic Manipulation of Vascular Development

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Rosa26-CreERT2 mice were purchased from the Jackson Laboratory (Stock No. 008463). VEGFR3 (Flt4) floxed mice47 (link), Vegfc floxed mice47 (link), Cdh5(PAC)-CreERT2 mice29 (link), membrane-retained VE-cadherin (Cdh5α) mice48 (link) and VE-cadherin (Cdh5) floxed49 (link) were previously described. Mice were bred according to standard protocols and maintained on a C57BL/6 or mixed background. Mating pairs were set up in the afternoon and vaginal plugs checked in the morning. For E18.5 embryo collection, tamoxifen (200μl at 25mg/ml dissolved in corn oil) was administered by oral gavage to pregnant dams on E14.5, E15.5, and E16.5 and medroxyprogesterone (100μl at 15mg/ml dissolved in DMSO) was administered by subcutaneous injection on E17.5 to prevent parturition. For E14.5 embryo collection, tamoxifen (200μl at 25mg/ml dissolved in corn oil) was administered by oral gavage to pregnant dams on either E9.5, E10.5, and E11.5 (embryos in Figure 2 and Supplemental Figure 1) or E10.5, E11.5, and E12.5 (embryos in Figures 4, 5, and 6 and Supplemental Figures 4 and 5). Cre-negative and/or heterozygous littermates were used as controls. All procedures were conducted using an approved animal protocol (806811) in accordance with the University of Pennsylvania Institutional Animal Care and Use Committee.
Sung D.C., Chen M., Dominguez M.H., Mahadevan A., Chen X., Yang J., Gao S., Ren A.A., Tang A.T., Mericko P., Patton R., Lee M., Jannaway M., Nottebaum A., Vestweber D., Scallan J.P, & Kahn M.L. (2022). Sinusoidal and lymphatic vessel growth is controlled by reciprocal VEGF-C–CDH5 inhibition. Nature cardiovascular research, 1(11), 1006-1021.
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10

Conditional Knockout Mice Generation

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B6, B6/SJL (CD45.1+ congenic) and Rag2–/–Il2rg–/– mice were from Taconic. Mettl3fl/fl mice were generated by J. H. Hanna at Weizmann Institute of Science. Klrg1Cre mice were generated and provided by R. Flavell at Yale University. Rorc-egfp reporter mice were provided by I. Ivanov at Columbia University. Rosa26CreERT2 mice were from Jackson Laboratories. To induce Cre expression in Mettl3Δ/Δ mice, the recipients were given 2 μg of tamoxifen in 100 μl of corn oil i.p. every other day for three doses. Mice used for experiments were females between 6 and 18 weeks of age, unless otherwise specifically indicated. All animal experiments were performed under the approval by the Institute Animal Care and Use Committee of Columbia University.
Minion F.C., Adams C, & Hsu T. (2000). R1 Region of P97 Mediates Adherence of Mycoplasma hyopneumoniae to Swine Cilia. Infection and Immunity, 68(5), 3056-3060.
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