B6n 129s1 tlr3tm1flv j

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The B6N.129S1-Tlr3tm1Flv/J mouse strain is a genetically modified model that carries a targeted mutation in the Tlr3 gene, which encodes the Toll-like receptor 3 (TLR3). TLR3 is a pattern recognition receptor that plays a role in the immune response to viral infections. This strain can be used in research related to the study of TLR3 function and its involvement in immune processes.

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4 protocols using b6n 129s1 tlr3tm1flv j

Wound Healing and Hair Follicle Regeneration

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All animal protocols are approved by the Johns Hopkins University Animal Care and Use Committee. C57BL/6J, B6;129SF2/J, TLR3 null mice (B6;129S1-Tlr3^tm1Flv/J and B6N.129S1-Tlr3tm1Flv/J), Nod-Scid-Gamma (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ), IL-6Rα^fl/fl (B6;SJL-Il6ra^tm1.1Drew/J) were obtained from The Jackson Laboratory. K5-Ert2-Cre and K14-Ert2-Cre mice were provided by Pierre Chambon. Stat3^fl/fl mice were kindly provided by Cynthia Sears (Johns Hopkins University; Stat3^tm2Aki(Takeda et al., 1998 (link))) and mixed strain (C57BL/6J x FVB/N x SJL/J) animals were provided by Dr. Jean Richa (University of Pennsylvania).
A 1 cm² excisional full-thickness wound to the level of skeletal muscle on the backs of 21-day old male and female mice was performed as previously described (Ito et al., 2007 (link); Nelson et al., 2013 (link)). Numbers of regenerated hair follicles were quantified in the re-epithelialized skin by non-invasive confocal scanning laser microscopy (CSLM) as published (Fan et al., 2011 (link)). For all experiments, 50μL of “intervention” was injected into healing wound (under scab) or applied topically into open wound as shown in the Table 1.

Nelson A.M., Reddy S.K., Ratliff T.S., Hossain M.Z., Katseff A.S., Zhu A.S., Chang E., Resnik S.R., Page C., Kim D., Whittam A.J., Miller L.S, & Garza L.A. (2015). dsRNA Released by Tissue Damage Activates TLR3 to Drive Skin Regeneration. Cell stem cell, 17(2), 139-151.

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Generation of Tamoxifen-Inducible HOIP-Deficient Mice

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Tlr3^−/− (9675, B6N.129S1-Tlr3^tm1Flv/J), cpdm (7599, C57BL/KaLawRij-Sharpin^cpdm/RijSunJ), and Tamoxifen-inducible Cre mice (4682, B6.Cg-Tg(CAG-cre/Esr1*)5Amc/J) were obtained from The Jackson Laboratory. HOIP-floxed mice were generated as previously described (Peltzer et al., 2014 (link)). HOIP-floxed mice were crossed with transgenic mice expressing the Tamoxifen-inducible loxP-deleter Cre recombinase to generate Tamoxifen-inducible HOIP-deficient mice. All mice were crossed for at least five generations before the experimental studies. All mice were typed by PCR analysis. Colonies were fed ad libitum. All animal experiments were conducted under an appropriate UK project license in accordance with the regulations of UK home office for animal welfare according to the ASPA (Animals [Scientific Procedures] Act 1986).

Zinngrebe J., Rieser E., Taraborrelli L., Peltzer N., Hartwig T., Ren H., Kovács I., Endres C., Draber P., Darding M., von Karstedt S., Lemke J., Dome B., Bergmann M., Ferguson B.J, & Walczak H. (2016). LUBAC deficiency perturbs TLR3 signaling to cause immunodeficiency and autoinflammation. The Journal of Experimental Medicine, 213(12), 2671-2689.

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Huntington's Disease Mice Tolerability

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B6C3-Tg(HD82Gln)81Dbo/J (N171-82Q) HD model mice, congenic TLR2^−/− mice (B6.129-Tlr2^tm1Kir/J, TLR3−/− mice (B6N.129s1-Tlr3tm1Flv/J) and TLR4^−/− mice (B6.B10ScN-Tlr4^lps−del/JthJ) were purchased from Jackson Laboratories (Bar Harbor, ME, USA). HD mice heterozygous for TLRs were generated by crossing HD mice with homozygous TLR2, TLR3 or TLR4 null mice. This study was conducted under the National Institute on Aging’s Institutional Animal Care and Use Committee (NIH animal welfare assurance number is: A4149-01). In accordance with these guidelines, mice were euthanized if they lost more than 20% of their initial body weight. When the animal exhibited clear neurological signs (unsteady gait, tremors/shakiness, lack of grooming) the animals were monitored on a daily basis by the caretakers and investigator in consultation with the attending veterinarian. to prevent dehydration in the animal due to a possible lack of food/water intake, Fruit jello or water soaked food, was placed on the floor of the cage, including daily injection of warm sterile saline (1.5–2 ml, intraperitoneally) if necessary. Mice were euthanized promptly when they were unable to ingest food or water. The aim of this study was to assess effects of genetic crosses on life expectancy, an experimental endpoint that was established at the beginning of the experiment.

Griffioen K., Mattson M.P, & Okun E. (2018). Deficiency of Toll-like receptors 2, 3 or 4 extends life expectancy in Huntington’s disease mice. Heliyon, 4(1), e00508.

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Murine Models for Influenza Research

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C57BL/6J and CD45.1⁺ coisogenic B6 mice were purchased from Jackson Laboratories and bred at the Heinrich Pette Institute animal facility. TLR3^-/- mice (B6N.129S1-Tlr3tm1Flv/J), OT-I mice (C57BL/6-Tg(TcraTcrb)1100Mjb/J), and Langerin-DTR mice (B6.129S2-Cd207tm3Mal/J) were purchased from Jackson Laboratories. All the experiments described were performed with males between 8-10 weeks of age unless otherwise stated. Animal experiments were conducted according to the guidelines of the German animal protection law. All staff carrying out animal experiments passed training programs according to category B or C of the Federation of European Laboratory Animal Science Associations. Experimental vaccination with cold-adapted LAIV as well as viral challenge was achieved by inoculation of the virus solution in PBS directly to the nostrils of mice anesthetized with Isoflurane. Poly IC (Invivogen) was administered intranasally at the indicated time points and concentrations in a PBS solution. Influenza A/PR8/34 (H1N1), X-31 (H3N2), and PR8-GFP viruses were propagated in 10-day-old embryonated chicken eggs at 37 °C.

Pérez-Girón J.V., Belicha-Villanueva A., Hassan E., Gómez-Medina S., Cruz J.L., Lüdtke A., Ruibal P., Albrecht R.A., García-Sastre A, & Muñoz-Fontela C. (2014). Mucosal poly IC improves protection elicited by replicating influenza vaccines via enhanced dendritic cell function and T cell immunity. Journal of immunology (Baltimore, Md. : 1950), 193(3), 1324-1332.

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