R26creert2

Manufactured by Jackson ImmunoResearch

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The R26CreERT2 is a Cre recombinase-expressing mouse strain. The Cre recombinase is fused to a mutated estrogen receptor (ERT2) that is responsive to the synthetic ligand 4-hydroxytamoxifen (4-OHT). This allows for temporal control of Cre-mediated recombination.

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6 protocols using r26creert2

Genetic Manipulation of Murine Immune Cells

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All mouse strains were bred and housed in specific pathogen-free conditions in accordance with the Institutional Animal Care and Use Guidelines of the University of California, San Diego (UCSD) at a temperature between 18 °C and 23 °C with 40–60% humidity. Male and female mice were both used in the present study. All mice used were on a C57BL/6J background. P14, Tgfbr2^fl/fl mice (stock no. 012603, Jackson Laboratory), R26Cre-ERT2 (stock no. 008463, Jackson Laboratory), Thy1.1 and CD45.1 congenic mice were bred in house. Prdm1^fl/fl (stock no. 008100, Jackson Laboratory) and Gzmb-cre (stock no. 003734, Jackson Laboratory) spleens were a gift from the laboratory of S. Kaech. To delete floxed alleles using Cre-ER^T2, we administered 1 mg of tamoxifen (Cayman Chemical Company) emulsified in 100 μl of sunflower seed oil (Sigma-Aldrich) via daily intraperitoneal injections on days 14–18 of infection. All animal studies were approved by the Institutional Animal Care and Use Committees of UCSD and performed in accordance with UC guidelines.

Basham B., Schroth G.P, & Ho P.S. (1995). An A-DNA triplet code: thermodynamic rules for predicting A- and B-DNA.. Proceedings of the National Academy of Sciences of the United States of America, 92(14), 6464-6468.

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Genetically Modified Mouse Lines for Immunology

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C57BL/6 (B6), CD45.1⁺ B6, R26^CreERT2, Irf4^flox/flox, Thy1.1⁺ Pmel-1 TCR-transgenic, and Tcf7^GFP flox (referred to as TCF1^GFP) mice were acquired from the Jackson Laboratory (Bar Harbor, ME). Irf4^−/− mice have been documented in prior studies [33 ]. We designed the Irf4^GFP-DTR mice and enlisted the assistance of Jackson Laboratory Model Generation Services to create this mouse line through the utilization of the CRISPR/Cas9 methodology [34 (link)]. We conducted a cross between Irf4^GFP-DTR and Thy1.1⁺ Pmel-1 mice to generate Irf4^GFP-DTR Thy1.1⁺ Pmel-1 mice. R26^CreERT2 mice were crossed to Irf4^flox/flox mice and then to Pmel-1 mice to generate R26^CreERT2Irf4^fl/fl CD45.2⁺ Pmel-1 mice. TCF1^GFP and Thy1.1⁺ Pmel-1 mice were crossed to generate TCF1^GFP Thy1.1⁺ Pmel-1 mice. Irf4^−/− and TCF1^GFP Thy1.1⁺ Pmel-1 mice were crossed to generate Irf4^−/−TCF1^GFP Thy1.1⁺ Pmel-1 mice. The Institutional Animal Care and Use Committee (IACUC) at Houston Methodist Research Institute granted approval for all animal-related procedures conducted in this study.

Yu A., Fu J., Yin Z., Yan H., Xiao X., Zou D., Zhang X., Zu X., Li X.C, & Chen W. (2023). Continuous Expression of Interferon Regulatory Factor 4 Sustains CD8+ T Cell Immunity against Tumor. Research, 6, 0271.

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Genetically-engineered Mouse Models

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Kras^G12DLSL/+, Kras^G12DFSF/+, R26^CreERT2 and R26^Cag-LSL-Luc mice were generated by T. Jacks and obtained through the Jackson Laboratory^{30 (link)–33 (link)}. Tp53^Frt/Frt mice were generated by D. Kirsch and obtained through the Jackson Laboratory^{34 (link)}. The R26^mTmG strain was generated by L. Luo and obtained through the Jackson Laboratory^{35 (link)}. The Smarcb1^Loxp/LoxP strain was provided by C. Roberts^{36 (link)}. The Pdx1-Cre strain was obtained from A. M. Lowy through the Jackson Laboratory^{37 (link)}. The Ptf1a^Cre/+ and Tp53^LoxP/LoxP strains were provided by R.A.D.^{38 (link),39 (link)}. The R26^Cag-FlpoERT2 was generated by A. Joyner and obtained from the Jackson Laboratory^{40 (link)}. The Cdh1^Cfp strain was generated by H. Clevers and obteined through the Jackson Laboratory^{41 (link)}. The Kras^G12DFSF/+; Tp53^Frt/Frt; R26^CreERT2 were kept in a C57BL/6 background, the other strains were kept in a mixed C57BL/6 and 129Sv/Jae background. All animal studies and procedures were approved by the UTMDACC Institutional Animal Care and Use Committee. Animals were killed when sick or when they developed tumours larger than 15 mm in their greater diameter or ulcerated lesions.

Genovese G., Carugo A., Tepper J., Robinson F.S., Li L., Svelto M., Nezi L., Corti D., Minelli R., Pettazzoni P., Gutschner T., Wu C.C., Seth S., Akdemir K.C., Leo E., Amin S., Molin M.D., Ying H., Kwong L.N., Colla S., Takahashi K., Ghosh P., Giuliani V., Muller F., Dey P., Jiang S., Garvey J., Liu C.G., Zhang J., Heffernan T.P., Toniatti C., Fleming J.B., Goggins M.G., Wood L.D., Sgambato A., Agaimy A., Maitra A., Roberts C.W., Wang H., Viale A., DePinho R.A., Draetta G.F, & Chin L. (2017). Synthetic vulnerabilities of mesenchymal subpopulations in pancreatic cancer. Nature, 542(7641), 362-366.

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Genetically Modified Mouse Models

Cited 1 time

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Wild-type C57BL6 mice were used unless specified otherwise. Runx3^−/−, TrkC^CreER, Raldh2^−/−, Raldh1^fl/fl, Raldh2^fl/fl, Raldh3^fl/fl and the CAGG^CreERTM mouse strains have been described elsewhere^{30 (link),48 (link),62 (link)–65 (link)}. Bax^−/−, R26^CreERT2, TrkC^−/− and Ai14 mice have been purchased from Jackson Laboratories, and NT3^−/− and TrkC^Cre from MMRRC. Animals of either sex were included in this study. Animals were group-housed, with food and water ad libitum, under 12 h light–dark cycle conditions.
Fertile white Leghorn eggs were incubated at 38 °C and embryos were staged according to Hamburger–Hamilton (HH) tables.
All animal work was performed in accordance with the national guidelines and approved by the local ethics committee of Stockholm, Stockholms Norra djurförsöksetiska nämnd.

Wang Y., Wu H., Fontanet P., Codeluppi S., Akkuratova N., Petitpré C., Xue-Franzén Y., Niederreither K., Sharma A., Da Silva F., Comai G., Agirman G., Palumberi D., Linnarsson S., Adameyko I., Moqrich A., Schedl A., La Manno G., Hadjab S, & Lallemend F. (2019). A cell fitness selection model for neuronal survival during development. Nature Communications, 10, 4137.

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Genetically Modified Mouse Strains

Cited 1 time

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CD-1 (strain 022) and FVB (strain 207) mice were purchased from Charles River Laboratories. CX3CR1^GFP (stock no. 005582), rd10 (stock no. 004297), C3H (stock no. 000659), sighted C3H (stock no. 003648), R26-CreERT2 (stock no. 008463), TSP1^–/– (stock no. 006141), and sighted FVB (stock no. 004828) mice were purchased from The Jackson Laboratory. Rho^–/– mice were a gift from Janis Lem (Tufts University, Boston, Massachusetts, USA) (46 (link)). SIRPα^–/– mice were a gift from Beth Stevens (Harvard Medical School) (26 (link)). CX3CR1^GFP, R26-CreERT2, TSP1^–/–, and SIRPα^–/– lines were crossed with FVB mice for at least 4 generations to obtain the following strains: sighted CX3CR1^GFP/+, rd1;CX3CR1^GFP/+, rd1;CreERT2/+, rd1;TSP1^+/-, rd1;TSP1^–/–, rd1;SIRPα^+/-, and rd1;SIRPα^–/–. Genotyping was performed by Transnetyx using real-time PCR. Mice were maintained at Harvard Medical School on a 12-hour alternating light and dark cycle. Animals were housed in standard ventilated racks at a density of up to 5 per cage. Both male and female animals were used in all experiments and were randomly assigned to experimental groups.

Wang S.K., Xue Y, & Cepko C.L. (2021). Augmentation of CD47/SIRPα signaling protects cones in genetic models of retinal degeneration. JCI Insight, 6(16), e150796.

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Characterization of Cdk8 Knockout Mice

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C57BL/6J mice and B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J (strain number 008463) hereinafter referred to as R26-Cre-ER^T2, were purchased from the Jackson Lab. Cdk8^fl/fl mice were kindly provided by Genentech and initially described by the Firestain group [6 (link)]. Animals were maintained under controlled room conditions (22–24 °C and a 14 h light:10 h dark photoperiod). Mice were given ad libitum access to food and water. Approximately two months old mice were used in the experiments. All manipulations with animals were performed according to the Local Bioethical Committee recommendations and according to the Declaration of Helsinki (1996).
To achieve the pregnancies, female mice were crossed overnight with males. The date of pregnancy was determined by observation of the vaginal plug.

Ilchuk L.A., Stavskaya N.I., Varlamova E.A., Khamidullina A.I., Tatarskiy V.V., Mogila V.A., Kolbutova K.B., Bogdan S.A., Sheremetov A.M., Baulin A.N., Filatova I.A., Silaeva Y.Y., Filatov M.A, & Bruter A.V. (2022). Limitations of Tamoxifen Application for In Vivo Genome Editing Using Cre/ERT2 System. International Journal of Molecular Sciences, 23(22), 14077.

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