Ifnar1 mice

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Ifnar1−/− mice are genetically modified mice that have a disrupted Ifnar1 gene, which encodes the type I interferon receptor subunit 1. These mice lack a functional type I interferon receptor and are commonly used in research to study the role of type I interferons in various biological processes.

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19 protocols using ifnar1 mice

In Vivo Evaluation of ZIKV Antibody Therapy

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Five-week-old type I interferon receptor-deficient (Ifnar1^−/−) mice with a C57BL/6J background were purchased from the Jackson Laboratory (Bar Harbor, ME, USA) and used to evaluate the in vivo efficacy levels of the ZV54 variants. As previously described [44 (link)], the mice were divided into two groups (n = 10 per group) and inoculated subcutaneously with 1 × 10⁵ PFUs of ZIKV on the ventral sides of their right hind footpads. Twenty-four hours post-infection, the mice in group 1 were intraperitoneally treated with 280 µg of pZV54^WT mAb and the mice in group 2 (control group) received an equivalent volume of PBS. Blood was collected in Trizol on day 3 post-infection to quantitate the level of viremia by RT-qPCR. The survival of the mice was monitored daily for 20 days post-ZIKV infection.

Sun H., Yang M., Lai H., Neupane B., Teh A.Y., Jugler C., Ma J.K., Steinkellner H., Bai F, & Chen Q. (2023). A Dual-Approach Strategy to Optimize the Safety and Efficacy of Anti-Zika Virus Monoclonal Antibody Therapeutics. Viruses, 15(5), 1156.

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Generating Knockout Murine Macrophages

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C57BL/6 mice (6-8 weeks old) were from Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (Beijing). The interferon-α/β receptor 1 (IFNAR1)-deficient (Ifnar1^-/-) mice (6-8 weeks old) were obtained from Jackson Laboratory. All mice were bred and maintained under specific-pathogen-free conditions. All animal experiments were performed according to the National Institutes of Health Guide for the Care and Use of Laboratory Animals, with the approval of the Animals Care and Use Committees of the Institute of Laboratory Animal Sciences of Chinese Academy of Medical Sciences (ACUC-A01-2021-040).
Mouse peritoneal macrophages were obtained as previously described (8 (link), 25 (link)). The RAW264.7, A549 and HEK293T cell lines were obtained from American Type Culture Collection (ATCC) and cultured as required. We generated FBL-knockdown RAW264.7 cells by a CRISPR-Cas9 gene-editing system with short guide RNA sequence-containing plasmid targeting specific sequences in the genome (5’-GGAGGTCGAGGTCGAGGCGG-3’ and 5’-GCTGCCAGCTTGGAGCGGAA-3’). We used PCR followed by sequencing and immunoblotting to determine the knockdown efficiency. MAVS^-/- A549 cells and MDA5^-/- A549 cells were also generated by a CRISPR-Cas9 approach.

Li P., Liu Y., Song R., Zhao L., Yang J., Lu F, & Cao X. (2022). RNA 2’-O-Methyltransferase Fibrillarin Facilitates Virus Entry Into Macrophages Through Inhibiting Type I Interferon Response. Frontiers in Immunology, 13, 793582.

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Syngeneic HNSCC Tumor Model and Immunotherapy

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Syngeneic HNSCC cells were implanted subcutaneously at the neck of 6–8 weeks C57BL/6 or Ifnar1^−/− mice (Jackson Laboratory). Tumor volume was calculated as ½(length × width²). In the irradiation experiments, mice were irradiated with a single 20-Gy dose or three fractionated doses of 8-Gy when tumors reached ~200mm³. For magnetic resonance imaging (MRI), nanosatellite-conjugated with E7 peptides were administered subcutaneously at tail-base at the 50 μg iron/mouse prior to imaging. To test the vaccines, mock (PBS, 100 μl), 2′3′-cGAMP (50 μg/100 μl) (Cat#tlrl-nacga23-1, InvivoGen), peptides (18.5 nmol/100 μl), or 3 weekly doses of SatVax [2′3′ cGAMP (50ug) and peptide (18.5 nmol) conjugated with the nanosatellite/100 μl] was administered subcutaneously at tail base. Intraperitoneal injections of anti-PD-L1 (100 μg/100 μl) (clone B7H1, BioXCell) were performed on day 1 and 4 after each vaccination with a total of 6 doses. For vaccination in hosts with established tumors, Montanide-E6/E7 peptide emulsion (100 μl/mouse), SatVax or PBS-mock was administered at tail base when tumors reached ~200mm³. All animal work were done in accordance with and approved by the Institutional Animal Care & Use Committee (PRO00006591).

Tan Y.S., Sansanaphongpricha K., Xie Y., Donnelly C.R., Luo X., Heath B.R., Zhao X., Bellile E., Hu H., Chen H., Polverini P.J., Chen Q., Young S., Carey T.E., Nör J.E., Ferris R.L., Wolf G.T., Sun D, & Lei Y.L. (2018). Mitigating SOX2-potentiated immune escape of Head and Neck Squamous Cell Carcinoma with a STING-inducing nanosatellite vaccine. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(17), 4242-4255.

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Mouse Isolation for Pathogen-Free Research

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All cell isolation involving live mice was performed in accordance with the recommendations from the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and the Office of Laboratory Animal Welfare. Mouse studies were performed using protocols approved by the Institutional Animal Care and Use Committee (IACUC). All mice were housed and bred under specific pathogen-free conditions and in accordance with Michigan State University (PROTO202200127) IACUC guidelines. All mice were monitored and weighed regularly. C57BL6/J mice (# 000664) and Ifnar1−/− mice (# 028288) were purchased from The Jackson Laboratory.

Ankley L.M., Conner K.N., Vielma T.E., Thapa M, & Olive A.J. (2023). GSK3α/β restrains IFNγ-inducible costimulatory molecule expression in alveolar macrophages, limiting CD4+ T cell activation. bioRxiv.

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Murine cell differentiation protocol

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C57BL/6 male mice (6-8 weeks of age) and Ifnar1 −/− mice were from the Jackson Laboratory. All mice experiments were performed in accordance with guidelines from the University of California, Los Angeles, Institutional Animal Care and Use Committee. HEK293T and RAW264.7 cells were purchased from ATCC and cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. RXRα^fl/fl (WT) and LysM-Cre +/RXRα^fl/fl (Rxra−/−) mice bone marrows were overnight shipped from Dr Mercedes Ricote's Lab (Spain)^{13 (link)}. WT, Ifnar1 −/− and Rxra −/− BMMs were differentiated as described previously^{45 (link)}. WT and Rxra −/− F9 embryocarcinoma cells, as well as RXR-specific agonists AGN194204 and LG268 were obtained from Dr Peter Tontonoz's Lab (University of California, Los Angeles). HX531 was obtained from Dr Hiroyuki Kagechika's lab (Tokyo Medical and Dental University). 9cRA and 13cRA were purchased from Sigma-Aldrich. All compounds for in vitro treatment were solubilized in DMSO. PolyI:C and polydA:dT were from InvivoGen. Antibody against α-tubulin was from Sigma-Aldrich. Anti-VSV-G (P5D4) and anti-Oct1 (C-21) antibodies were from Santa Cruz Biotechnology. Anti-RXRα (D6G10), anti-β-Actin (13E5) and anti-β-Catenin (D10A8) were from Cell Signaling Technology. Anti-GAPDH (GT239) was from GeneTex.

Ma F., Liu S.Y., Razani B., Arora N., Li B., Kagechika H., Tontonoz P., Núñez V., Ricote M, & Cheng G. (2014). Retinoid X receptor α attenuates host antiviral response by suppressing type I interferon. Nature communications, 5, 5494.

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Genetically Modified Mouse Models

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C57BL/6 and IFNAR1^–/– mice were purchased from Jackson Laboratories (Bar Harbor, ME, United States). IFNAR1^–/–, IRF3^–/–, IRF7^–/–, and K1 RBC transgenic mice were previously described (37 (link)–39 (link)). K1 mice express the KEL glycoprotein, containing the KEL1 antigen, on RBCs. Appropriate gene-deficient mice were bred to produce IRF3/7^–/– double knockout mice. All mice were 8–12 weeks of age and had been backcrossed to the C57BL/6 background for more than 8 generations. All pristane-treated mice were female mice injected intraperitoneally with one dose of 0.5 mL pristane (2,6,10,14-tetramethylpentadecane). All animal protocols were approved by the Cedars-Sinai Institutional Animal Care and Use Committee.

Lee J.Y., Madany E., El Kadi N., Pandya S., Ng K., Yamashita M., Jefferies C.A, & Gibb D.R. (2020). Type 1 Interferon Gene Signature Promotes RBC Alloimmunization in a Lupus Mouse Model. Frontiers in Immunology, 11, 584254.

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USUV Infection in Ifnar1-/- Mice

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Interferon α/β receptor 1 knockout (Ifnar1^-/-) mice, on a C57BL/6 background, were originally received from Jackson Labs and bred on site. In two biologically independent experiments, groups of 8 (4 female and 4 male) 8-week-old mice were subcutaneously inoculated via rear footpad injection with 10³ PFU of USUV isolates. Blood samples were collected in serum separator tubes via submandibular vein bleeds on days 1, 3, 5 and 7 post-inoculation. Mice were weighed daily and observed for clinical signs of illness, including lethargy, tremors, and weight loss. When mice exhibited ≥15% weight loss (or at 28 days post inoculation), they were euthanized by deep isoflurane anesthesia followed by cervical dislocation. Brain, heart, liver, and spleen tissues were collected at the time of euthanasia. In a separate experiment, an additional 4 mice were inoculated with TM Netherlands and euthanized on dpi 6, in order to collect tissues for comparison to other strains. Tissues were weighed and suspended in equal parts BA-1 medium, then homogenized via bead homogenization in a Qiagen TissueLyserLT at 50 oscillations/sec for 2 minutes followed by centrifugation to pellet solids. All samples were titrated by Vero cell plaque assay.

Kuchinsky S.C., Hawks S.A., Mossel E.C., Coutermarsh-Ott S, & Duggal N.K. (2020). Differential pathogenesis of Usutu virus isolates in mice. PLoS Neglected Tropical Diseases, 14(10), e0008765.

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Mouse Cell Isolation Protocol

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All cell isolation involving live mice was performed in accordance with the recommendations from the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and the Office of Laboratory Animal Welfare. Mouse studies were performed using protocols approved by the Institutional Animal Care and Use Committee. All mice were housed and bred under specific pathogen-free conditions and in accordance with Michigan State University (PROTO202200127) Institutional Animal Care and Use Committee guidelines. All mice were monitored and weighed regularly. C57BL6/J mice (catalog no. 000664) and Ifnar1^−/− mice (catalog no. 028288) were purchased from The Jackson Laboratory.

Ankley L.M., Conner K.N., Vielma T.E., Godfrey J.J., Thapa M, & Olive A.J. (2024). GSK3α/β Restrain IFN-γ–Inducible Costimulatory Molecule Expression in Alveolar Macrophages, Limiting CD4+ T Cell Activation. ImmunoHorizons, 8(2), 147-162.

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STING1 Mutant Mouse Generation

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The Sting1^S365A/S365A mutant mouse was generated by CRISPR/Cas9 technology at UT Southwestern Transgenic Mouse Facility on the C57BL/6 background. Sting1^−/− mice (C57BL/6 background) were obtained from Glen Barber (Univ. of Miami). Wild type and Rag1^−/− mice were purchased from UTSW Mouse Breeding Core. Irf3^−/− mice were obtained from Kate Fitzgerald (Univ. of Mass Medical School) with permission from Tadatsugu Taniguchi. Ifnar1^−/− mice were purchased from Jackson Labs. Both male and female mice were used. Most studies used 6–8 week old mice. All mice were maintained in pathogen-free barrier facilities and were used in accordance with protocols approved by the Institutional Animal Care and Use Committee at University of Texas Southwestern Medical Center.

Wu J., Dobbs N., Yang K, & Yan N. (2020). Interferon-independent activities of mammalian STING mediate antiviral response and tumor immune evasion. Immunity, 53(1), 115-126.e5.

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Mouse Influenza and Sendai Virus Protocols

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All general reagents were obtained from Sigma-Aldrich (St. Louis, MO), except as indicated. All mouse experiments were performed with 5- to 8-week-old C57BL/6J or Ifnar1^−/− mice (The Jackson Laboratory, Bar Harbor, ME) of a single gender, in accordance with the Institutional Animal Care and Use Committee of the University of Texas MD Anderson Cancer Center, protocol 00000907-RN01. Immortalized human bronchial epithelial (HBEC3kt) cells were kindly provided by John Minna. Murine lung epithelial (MLE-15) cells were kindly provided by Jeffrey Whitsett. Normal human bronchial epithelial (NHBE) cells were purchased from Lonza (Basel, Switzerland). Immortalized cells were authenticated by the MD Anderson Characterized Cell Line Core Facility. Mouse-adapted influenza A/Hong Kong/8/68 virus (H3N2; Mouse Lung Pool 1/17/12) was kindly provided by Brian E. Gilbert (58 (link)). Sendai virus (parainfluenza virus type 1) was obtained from the American Type Culture Collection (ATCC; Manassas, VA).

Kirkpatrick C.T., Wang Y., Leiva Juarez M.M., Shivshankar P., Pantaleón García J., Plumer A.K., Kulkarni V.V., Ware H.H., Gulraiz F., Chavez Cavasos M.A., Martinez Zayes G., Wali S., Rice A.P., Liu H., Tour J.M., Sikkema W.K., Cruz Solbes A.S., Youker K.A., Tuvim M.J., Dickey B.F, & Evans S.E. (2018). Inducible Lung Epithelial Resistance Requires Multisource Reactive Oxygen Species Generation To Protect against Viral Infections. mBio, 9(3), e00696-18.

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