B6 cg tlr4tm1.2karp j

Manufactured by Jackson ImmunoResearch

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The B6(Cg)-Tlr4tm1.2Karp/J is a strain of genetically modified mice. It has a targeted mutation in the Tlr4 gene, which is involved in the immune response. This strain can be used in research related to the Toll-like receptor 4 signaling pathway.

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7 protocols using b6 cg tlr4tm1.2karp j

Transgenic Mouse Strains for Research

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C57BL/6-Tg(Csf1r-HBEGF/mCherry)1Mnz/J (stock #024046), B6.129P2-Lyz2tm1(cre)Ifo/J (stock #004781), B6(Cg)-Tlr4tm1.2Karp/J (stock #029015), and B6.129S7-Rag1tm1Mom/J (stock #002216) were purchased from The Jackson Laboratory (Bar Harbor, ME) and used per vendor’s guidelines.

MohanKumar K., Namachivayam K., Song T., Jake Cha B., Slate A., Hendrickson J.E., Pan H., Wickline S.A., Oh J.Y., Patel R.P., He L., Torres B.A, & Maheshwari A. (2019). A murine neonatal model of necrotizing enterocolitis caused by anemia and red blood cell transfusions. Nature Communications, 10, 3494.

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Engineered Mouse Models for Research

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Animals were bred in-house by OHSU staff. C57BL/6 and Tlr4-KO (B6(Cg)-Tlr4^tm1.2Karp/J) founder mice were procured from Jackson Lab. G-csf-KO (B6.129X1(Cg-Csf3r^tm1Link/J) founder mice were generously provided by Dr. Jason Shohet. Both mutant strains have a C57BL/6 background. Both male and female mice were used at ages 16–20 weeks with weights ranging from 18–30 g. Animals were housed in gender-separated boxes in groups of between 3–6 mice. Animals were allowed free access to food and water and kept under a twelve-hour light/dark cycle.

Urdang Z.D., Bills J.L., Cahana D.Y., Muldoon L.M, & Neuwelt E.A. (2020). Toll-like Receptor 4 Signaling and Downstream Neutrophilic Inflammation Mediate Endotoxemia-Enhanced Blood-Labyrinth Barrier Trafficking. Otology & neurotology : official publication of the American Otological Society, American Neurotology Society [and] European Academy of Otology and Neurotology, 41(1), 123-132.

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Bone Marrow Mast Cell Isolation

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For experiments involving Tlr4^−/− BMMCs, B6(Cg)-Tlr4^tm1.2Karp/J (#029015) and corresponding C57BL/6J WT controls (#000664), 8 weeks old female mice were purchased from The Jackson Laboratory. For all other experiments involving bone marrow or BMMCs, C57BL/6 WT mice (8–14 weeks old; male and female), bred and maintained at the National Veterinary Institute (SVA, Uppsala, Sweden) were used. The experimental procedures were approved by the local animal ethics committee (Uppsala djurförsöksetiska nämnd, Dnr 5.8.18-05357/2018). Whenever possible, remaining bones from WT C57BL/6 mice used as controls in other experiments were acquired for culturing BMMCs.

von Beek C., Fahlgren A., Geiser P., Di Martino M.L., Lindahl O., Prensa G.I., Mendez-Enriquez E., Eriksson J., Hallgren J., Fällman M., Pejler G, & Sellin M.E. (2024). A two-step activation mechanism enables mast cells to differentiate their response between extracellular and invasive enterobacterial infection. Nature Communications, 15, 904.

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Genetically Modified Rodent Models

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Sprague–Dawley (SD) rats and C57BL/6 were purchased from ORIENT BIO. B6.129-Tlr2^tm1Kir/J, B6(Cg)-Tlr4^tm1.2Karp/J, and B6N.129S-Havcr2^tm1Bmed/J (Tim-3^mut) were purchased from The Jackson Laboratory. Mice carrying HIF1α-floxed alleles (HIF1α^+f/+f) were obtained from Dr. Randall Johnson (University of Cambridge, Cambridge, United Kingdom), and mice lacking HIF1α in myeloid lineage cells were generated by crossbreeding HIF1α^+f/+f mice to LysM-Cre transgenic mice. Mice carrying gene targeting of the Rosa26 locus with a Flag-TIM-3 construct (FSF-TIM-3) were produced by Dr. Lawrence P. Kane (University of Pittsburgh, Pittsburgh, PA; ref. 9). All animals were maintained and bred under SPF conditions in the Association for Assessment and Accreditation of Laboratory Animal Care–accredited National Cancer Center animal facility. All animal procedures were performed according to ARRIVE guidelines and NCC guidelines for the care and use of laboratory animals. The protocol was approved by the Committee on the Ethics of Animal Experiments of the NCC (Permit No.: NCC-11-125). To avoid bias, the animal studies in this study were properly randomized in a blinded manner with respect to the genotypes and treatments.

Kim H.S., Chang C.Y., Yoon H.J., Kim K.S., Koh H.S., Kim S.S., Lee S.J., Kane L.P, & Park E.J. (2020). Glial TIM-3 Modulates Immune Responses in the Brain Tumor Microenvironment. Cancer research, 80(9), 1833-1845.

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Murine Models for Immunological Studies

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C57BL/6 (WT), B6.SJL-Ptprc^aPepc^b/BoyJ, C57BL/6-Tg(UBC-GFP)30Scha/J (GFP⁺), B6.129P2(SJL)-Myd88^tm1.1Defr/J, B6.129-Tlr2^tm1Kir/J, B6(Cg)-Tlr4^tm1.2Karp/J, B6.129S1-Tlr7^tm1Flv/J, C57BL/6J-Tlr9^M7Btlr/Mmjax, B6.129S7-Il1r1^tm1/mx/J, C57BL/6J-Ticam1^Lps2/J, B6.Cg-Ccl2^tm1.1Pame/J, and B6.129S4-Ccr2^tm1lfc/J (The Jackson Laboratory) were used in this study. Csf2^−/− and Csf2rb^−/− mice on C57BL/6 background (10 generations) were bred in-house (Rauch et al., 2012 (link); Hilgendorf et al., 2014b (link); Weber et al., 2014 (link)). All experiments were conducted with male mice aged 8–16 wk at the time of death. All protocols were approved by the Animal Review Committee at Massachusetts General Hospital.

Anzai A., Choi J.L., He S., Fenn A.M., Nairz M., Rattik S., McAlpine C.S., Mindur J.E., Chan C.T., Iwamoto Y., Tricot B., Wojtkiewicz G.R., Weissleder R., Libby P., Nahrendorf M., Stone J.R., Becher B, & Swirski F.K. (2017). The infarcted myocardium solicits GM-CSF for the detrimental oversupply of inflammatory leukocytes. The Journal of Experimental Medicine, 214(11), 3293-3310.

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Investigating TLR4 Knockout Mice

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Inbred male wild-type (WT) C57Bl/6J and TLR4 knockout mice on a C57Bl/6J background (B6(Cg)-Tlr4tm1.2Karp/J, 029015), weighing 20–25 g were purchased from the Jackson Laboratory (Bar Harbor, ME) and were housed under pathogen-free conditions at the University of Colorado Denver, Barbara Davis Center Animal Facility according to NIH Guidelines and with approval of the University of Colorado Denver IACUC. Experiments were performed at the Transplant Microsurgery Facility (University of Colorado). The study protocol was approved by the University of Colorado Denver Institutional Animal Care and Use Committee. The study was carried out in compliance with the ARRIVE guidelines.

Jain S., Plenter R., Nydam T., Gill R.G, & Jani A. (2021). Deletion of TLR4 reduces apoptosis and improves histology in a murine kidney transplant model. Scientific Reports, 11, 16182.

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Burn Injury Model in Transgenic Mice

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Female C57BL/6J, B6.129P2(SJL)-Myd88^tm1.1Defr/J, B6(Cg)-Tlr4^tm1.2Karp/J, and B6.129-Tlr2^tm1Kir/J were obtained from Jackson Laboratory (Bar Harbor, ME) and used in studies at 8–12 weeks of age. A flame burn injury model was utilized to produce a full-thickness insensate lesion encompassing 15% of total body surface area, as previously described (27 ). In brief, mice were anesthetized with isoflurane and covered with a flame-resistant template exposing shaved dorsal skin. The target area was saturated with 0.5 ml of absolute ethanol and ignited for 10 seconds in the burn group or allowed to evaporate without ignition in the sham group. Immediately after the procedure, all mice received 0.5 ml of saline for volume resuscitation via the intraperitoneal (i.p.) route. Mice were sacrificed by i.p. injection of Euthasol (Virbac, Fort Worth, TX). All animal procedures were approved by the University of Cincinnati Institutional Care and Use Committee.

Nicolas F.J., Cayuela M.L., Martínez-Argudo I.M., Ruiz-Vazquez R.M, & Murillo F.J. (1996). High mobility group I(Y)-like DNA-binding domains on a bacterial transcription factor.. Proceedings of the National Academy of Sciences of the United States of America, 93(14), 6881-6885.

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