Tlr7 mice

Manufactured by Jackson ImmunoResearch

Sourced in Montenegro, United States

Tlr7−/− mice are genetically modified mice that have a deficiency in the Toll-like receptor 7 (Tlr7) gene. These mice lack the expression of the Tlr7 receptor, which plays a role in the recognition of single-stranded RNA and the subsequent immune response. The Tlr7−/− mice can be used in research to study the functions and importance of the Tlr7 receptor in various biological processes.

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Lab products found in correlation

C57bl 6, by Jackson ImmunoResearch (2 mentions) Isopro rmh 3000, by LabDiet (1 mentions) Batf3 mice, by Jackson ImmunoResearch (1 mentions) Balb c mice, by Jackson ImmunoResearch (1 mentions) C57bl 6j mice, by Jackson ImmunoResearch (1 mentions) Vetbond, by 3M (1 mentions) Csfr1cre mice, by Jackson ImmunoResearch (1 mentions) Wt c57bl 6n mice, by Charles River Laboratories (1 mentions) Normocin, by InvivoGen (1 mentions) Zeocin, by InvivoGen (1 mentions) Blasticidin, by InvivoGen (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Sprague dawley rat, by Charles River Laboratories (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Raw264.7 macrophage, by American Type Culture Collection (1 mentions)

9 protocols using tlr7 mice

Genetically Modified Mouse Strains

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Wild-type (WT, C57BL/6), TLR3^-/-, and TLR7^-/- mice were purchased from The Jackson Laboratory (Bar
Harbor, ME). MyD88^-/- mice were generated by Kawai and colleagues (18 (link)).
Trif^-/- mice were generated by Yamamoto, et al. (19 (link)). Mice
were 8-12 week-old, weighed between 20-30 g, and gender and age matched. Mice were fed the same bacteria-free diet (Prolab Isopro
RMH 3000, LabDiet, Brentwood, MO) and water. The animal protocols were approved by the Subcommittee on Research Animal Care of
Massachusetts General Hospital (Boston, MA). The experiments were performed in compliance with the guideline of the National
Institutes of Health (Bethesda, MD). Simple randomization method was used to assign animals to various experimental
conditions.

Feng Y., Zou L., Yan D., Chen H., Xu G., Jian W., Cui P, & Chao W. (2017). Extracellular miRNAs induce potent innate immune responses via TLR7/MyD88-dependent mechanisms. Journal of immunology (Baltimore, Md. : 1950), 199(6), 2106-2117.

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In Vivo Mouse Experiments

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All in vivo experiments were performed in the following mouse strains: 8-week-old female C57BL/6J mice (the Jackson Laboratory), 8-week-old female BALB/c mice (the Jackson Laboratory), 8-week-old BATF3⁻ mice (the Jackson Laboratory, strain no. 013755), and 8-week-old TLR7⁻ mice (the Jackson Laboratory, strain no. 008380). Experiments were performed in specific pathogen–free animal facilities at the MIT Koch Institute for Integrative Cancer Research. Mice were housed under standard 12-hour light–12-hour dark conditions with ad libitum access to water and chow. All mouse studies were performed according to institutional and National Institutes of Health guidelines for humane animal use and in accordance with the Association for Assessment and Accreditation of Laboratory Animal Care. Protocols were approved by the Institutional Animal Care and Use Committee at MIT.

Bhagchandani S.H., Vohidov F., Milling L.E., Tong E.Y., Brown C.M., Ramseier M.L., Liu B., Fessenden T.B., Nguyen H.V., Kiel G.R., Won L., Langer R.S., Spranger S., Shalek A.K., Irvine D.J, & Johnson J.A. (2023). Engineering kinetics of TLR7/8 agonist release from bottlebrush prodrugs enables tumor-focused immune stimulation. Science Advances, 9(16), eadg2239.

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Partial Infraorbital Nerve Ligation Model

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ICR male mice weighing 26 g–30 g were provided by the Experimental Animal Center at Nantong University. Tlr8^−/− mice were developed by Cyagen Co. (Suzhou, China). Tlr7^−/− mice were purchased from the Jackson Laboratory (stock number 008380). All mice were housed in standard clear plastic cages under controlled ambient temperature (22 °C–24 °C) with a reversed 12:12 h dark/light cycle, and allowed ad libitum access to water and food. The experimental and surgical procedures were reviewed and approved by the Animal Care and Use Committee of Nantong University. Animal treatments were performed in accordance with the guidelines of the International Association for the Study of Pain.
The pIONL surgery was as described by Zhang et al. [24 (link), 25 (link)]. In brief, mice were anesthetized with sodium pentobarbital (40 mg/kg–50 mg/kg, i.p.), and then the oral cavity was opened to locate the tendon of the left masseter muscle on the upper wall of oral cavity in the supine position. In front of the tendon, an incision (1 mm) was made to expose the infraorbital nerve. The TNP model was established by ligating one-half of the infraorbital nerve (ION) with 6-0 silk suture. The mucous membrane of the incision was closed with Tissue Adhesive (Vetbond; 3M, USA). The sham operation comprised an incision on the mucous membrane without damaging the ION.

Zhao L.X., Jiang M., Bai X.Q., Cao D.L., Wu X.B., Zhang J., Guo J.S., Chen T.T., Wang J., Wu H., Gao Y.J, & Zhang Z.J. (2020). TLR8 in the Trigeminal Ganglion Contributes to the Maintenance of Trigeminal Neuropathic Pain in Mice. Neuroscience Bulletin, 37(4), 550-562.

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Generation and Characterization of Knockout Mouse Cell Lines

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Stocks of Nodamura virus (NoV) were produced by intraperitoneal injection of BALB/c suckling mice as previously reported [22 (link)]. We followed the guidelines described under the federal Animal Welfare Regulations Act with the protocol approved by the Institutional Animal Care and Use Committee at the University of California, Riverside. Mouse embryonic fibroblasts (MEFs) cell lines were generated from the wild-type (WT) mice with a C57BL/6 background and RIG-I^−/−, MDA5^−/−, LGP2^−/−, MAVS^−/−, TLR3^−/−, TLR7^−/− knockout mice as previously described [5 (link),10 (link),32 (link),33 (link),34 (link),35 (link)]. C57BL/6, MDA5^−/−, MAVS^−/−, TLR3^−/−, and TLR7^−/− mice were purchased from Jackson Laboratory whereas RIG-I^−/− and LGP2^−/− were kindly provided by Drs. Adolfo García-Sastre, Shizuo Akira and Michael Gale, Jr. Cells were maintained in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine solution, 37°C, 5% CO₂.

Fan X., Dong S., Li Y., Ding S.W, & Wang M. (2015). RIG-I-dependent antiviral immunity is effective against an RNA virus encoding a potent suppressor of RNAi. Biochemical and biophysical research communications, 460(4), 1035-1040.

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Mouse Experimental Protocol for TLR7 and EGFR

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All animal experiments were performed under the animal protocol (IACUC#: 007158) approved by the Institutional Animal Care & Use Committee at CSMC. Female 8–12 weeks mice were used for all experiments. Wild type (WT; C57Bl/6) and TLR7^−/− mice were purchased from The Jackson Laboratory (Bar Harbor, ME). EGFR^{floxed/floxed} mice were obtained from David Threadgill, PhD at Texas A&M University, and crossbred to CSFR1^Cre mice (The Jackson Laboratory) to generate EGFR^{floxed/floxed}- CSFR1^Cre mice. Mice were bred and kept in a specific pathogen-free condition with 12-h day/night cycles and allowed free access to food and water.

Lyons J, & Khot A. (2000). Managing information overload: developing an electronic directory. BMJ : British Medical Journal, 320(7228), 160.

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Murine Model of TLR7 Deficiency

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All animal procedures were performed in strict accordance with the protocols approved by the Institutional Animal Care and Use Committee at the University of Nebraska Medical Center (UNMC) and the NIH. C57BL/6N WT mice were purchased from Charles River Laboratories (Wilmington, MA, USA) and housed under conditions of constant temperature and humidity on a 12-hr light and 12-hr dark cycle, with lights on at 7:00 a.m. Food and water were available ad libitum. Tlr7^−/− mice were purchased from Jackson Laboratories (Bar Harbor, ME, USA) and bred in the UNMC animal facility. Pregnant WT mice were purchased from Charles River Laboratories.

Hu G., Liao K., Niu F., Yang L., Dallon B.W., Callen S., Tian C., Shu J., Cui J., Sun Z., Lyubchenko Y.L., Ka M., Chen X.M, & Buch S. (2018). Astrocyte EV-Induced lincRNA-Cox2 Regulates Microglial Phagocytosis: Implications for Morphine-Mediated Neurodegeneration. Molecular Therapy. Nucleic Acids, 13, 450-463.

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TLR7 Activation in Immune Cells

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miR-21^−/− and Tlr7^−/− mice were purchased from Jackson Laboratories (Bar Harbor, Maine) and bred in the UNMC animal facility. Pregnant WT mice were purchased from Charles River (Wilmington, MA, USA). HEK-Blue TLR7 cells designed for studying the stimulation of TLR7 by monitoring the activation of NF-κB and AP-1 were cultured in DMEM supplemented with 10% FBS, normocin (50 μg/ml), blasticidin (10 μg/ml), zeocin (100 μg/ml) (InvivoGen, San Diego, CA). Cells were grown at 37° C in humidified air with 5% CO₂. Control HEK-Blue Null cells were cultured similarly except without zeocin.

Yelamanchili S.V., Lamberty B.G., Rennard D.A., Morsey B.M., Hochfelder C.G., Meays B.M., Levy E, & Fox H.S. (2015). MiR-21 in Extracellular Vesicles Leads to Neurotoxicity via TLR7 Signaling in SIV Neurological Disease. PLoS Pathogens, 11(7), e1005032.

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Microglial Cell Lines and Knockout Mice

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Tlr7^−/− mice were purchased from Jackson Laboratories (Bar Harbor, Maine) and bred in the UNMC animal facility. Pregnant wildtype (WT) mice and Sprague Dawley rats were purchased from Charles River (Wilmington, MA, USA). RAW 264.7 macrophages (American Type Culture Collection, Manassus, VA, USA) were cultured in Dulbecco's modified eagle's medium (DMEM) (Invitrogen, CA, USA) supplemented with 10% fetal bovine serum (FBS) (Invitrogen) and penicillin‐streptomycin. The NR‐9460 microglial cell line, derived from wild‐type mice (WT‐MG), and the NR‐19980 microglial cell line, derived from toll‐like receptor 7 (TLR7) knockout mice (denoted TLR^‐/‐‐MG) (BEI Resources, Manassus, VA, USA), were cultured in DMEM supplemented with 10% FBS, 2 mM L‐glutamine, 1 mM sodium pyruvate and 10 μg/ml ciprofloxacin. Cells were grown to confluence at 37°C in humidified air with 5% CO₂. Details for the cell lines are provided in supplemental files S1&S2.

Chand S., Gowen A., Savine M., Moore D., Clark A., Huynh W., Wu N., Odegaard K., Weyrich L., Bevins R.A., Fox H.S., Pendyala G, & Yelamanchili S.V. (2021). A comprehensive study to delineate the role of an extracellular vesicle‐associated microRNA‐29a in chronic methamphetamine use disorder. Journal of Extracellular Vesicles, 10(14), e12177.

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Murine Cancer Immunotherapeutic Protocols

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BALB/c, C57BL/6 and SCID mice were purchased from the Shanghai SLAC Laboratory Animal Company, China. TLR7−/− mice were purchased from the Jackson Laboratory (Bar Harbor, ME). All animals were maintained under specific pathogen-free conditions in the animal facilities of Shanghai Jiaotong University, School of Medicine. The murine colon cancer cell line CT-26 and the Lewis lung carcinoma tumor cell line LLC were purchased from Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. Tumor cells were cultured in complete RPMI 1640 medium supplemented with 10% FBS, 100 U/ml penicillin and 100 g/ml streptomycin in a humidified cell incubator with an atmosphere of 5% CO₂ at 37°C. Exponentially growing cells were used for experiments. TLR7 ligand Loxoribin was purchased from Invivogene (San Diego, CA, USA).

Wang C., Zhou Q., Wang X., Wu X., Chen X., Li J., Zhu Z., Liu B, & Su L. (2014). The TLR7 agonist induces tumor regression both by promoting CD4+T cells proliferation and by reversing T regulatory cell-mediated suppression via dendritic cells. Oncotarget, 6(3), 1779-1789.

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