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Nod cg rag1tm1mom il2rgtm1wjl szj

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The NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ mouse is a laboratory animal model characterized by a deficiency in the recombination-activating gene 1 (Rag1) and the interleukin-2 receptor gamma chain (Il2rg) genes. This model is used in immunological research to study the effects of these genetic modifications.

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Rag1tm1Mom (13 citations)

20 protocols using nod cg rag1tm1mom il2rgtm1wjl szj

1

SARS-CoV-2 Infection in Humanized Mice

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NOD Rag1−/−IL2Rgnull mice (NOD.Cg-Rag1tm1MomIl2rgtm1Wjl/SzJ, were obtained from the Jackson Laboratory, catalog number 007799). NRG-Flk2−/− (NRGF) mice (NOD.Cg-Rag1tm1MomFlt3tm1IrlIl2rgtm1Wjl/J) were generated as described previously (Douam et al., 2018 (link)) and are available at The Jackson Laboratory (Bar Harbor, ME, USA) (catalog number 033127). NRG and NRGF mice were maintained at the Laboratory Animal Resource Center at Princeton University prior to engraftment with human tissues and shipment to the NEIDL.
In the NEIDL BSL-3 facility, mice were group-housed by sex in Tecniplast green line individually ventilated cages (Tecniplast, Buguggiate, Italy). Mice were maintained on a 12:12 light cycle at 30–70% humidity and provided sulfatrim-containing water and standard chow diets (LabDiet, St. Louis, MO, USA).
All mice in this study were inoculated with SARS-CoV-2 at an age of 20–30 weeks old. Both male and female mice were used.
Kenney D.J., O’Connell A.K., Turcinovic J., Montanaro P., Hekman R.M., Tamura T., Berneshawi A.R., Cafiero T.R., Al Abdullatif S., Blum B., Goldstein S.I., Heller B.L., Gertje H.P., Bullitt E., Trachtenberg A.J., Chavez E., Nono E.T., Morrison C., Tseng A.E., Sheikh A., Kurnick S., Grosz K., Bosmann M., Ericsson M., Huber B.R., Saeed M., Balazs A.B., Francis K.P., Klose A., Paragas N., Campbell J.D., Connor J.H., Emili A., Crossland N.A., Ploss A, & Douam F. (2022). Humanized mice reveal a macrophage-enriched gene signature defining human lung tissue protection during SARS-CoV-2 infection. Cell Reports, 39(3), 110714.
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2

Evaluating Tumor Immune Modulation in Xenograft Models

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All animal experiments were conducted in accordance with and the approval of the MSKCC IACUC (Institutional Animal Care and Use Committee) protocols. Female NSG (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ) and NRG (NOD.Cg-Rag1tm1MomIl2rgtm1Wjl/SzJ) mice were purchased from the Jackson Laboratory at 5-10 weeks old. For experiments in vivo with RET and ALK, 2.5-6 ×106 tumor cells in PBS were subcutaneously injected into the flank of mice. When tumors were palpable (2-3 mm), mice were treated daily with drugs or vehicles through oral gavage of drugs in 200ul of water. At day 4 or 7, tumors were harvested and flow cytometry was conducted to determine the effect of inhibitors on HLA and PD-L1 on the tumor cells. N=5 for all treatment groups in vivo (one outlier was excluded from the vehicle group in the RET experiment, but results remained significant: P value with outlier included 0.031, P value without outlier 0.016)). TPC1 cells were transduced with luciferase and GFP on an SFG vector and this allowed gating of the tumor cells in flow cytometry. A CD30 antibody was used in the ceritinib experiments.
Oh C.Y., Klatt M.G., Bourne C., Dao T., Dacek M.M., Brea E.J., Mun S.S., Chang A.Y., Korontsvit T, & Scheinberg D.A. (2019). ALK and RET inhibitors promote HLA class I antigen presentation and unmask new antigens within the tumor immunopeptidome. Cancer immunology research, 7(12), 1984-1997.
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3

Humanized Mouse Model for HIV-1 Research

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NOD Rag1−/− IL2RγNULL mice were humanized with hematopoietic stem cells (HSCs), as previously described (Klein et al., 2012 (link)). In brief, human fetal livers were procured from Advanced Bioscience Resources, Inc. and HSCs were isolated using a CD34+ isolation kit (Stem Cell Technologies, Inc.). Mice (NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ) were obtained from The Jackson Laboratory and bred and maintained at the Comparative Bioscience Center of The Rockefeller University. Between 1–5-d-old mice were irradiated with 100 cGy and injected with 2 × 105 human HSCs. HSCs used to reconstitute the mice were obtained from human fetal liver tissues of 12 different donors. In the experiments shown, up to 12 humanized mice shared the same donor. Engraftment was evaluated by FACS analysis of the peripheral blood as previously described (Klein et al., 2012 (link)) and mice with successful reconstitution of human lymphocytes were challenged with HIV-1YU2 or HIV-1YU2TM2. All experiments were performed under approval of the Institutional Review Board and the Institutional Animal Care and Use Committee of The Rockefeller University.
Klein F., Nogueira L., Nishimura Y., Phad G., West AP J.r., Halper-Stromberg A., Horwitz J.A., Gazumyan A., Liu C., Eisenreich T.R., Lehmann C., Fätkenheuer G., Williams C., Shingai M., Martin M.A., Bjorkman P.J., Seaman M.S., Zolla-Pazner S., Karlsson Hedestam G.B, & Nussenzweig M.C. (2014). Enhanced HIV-1 immunotherapy by commonly arising antibodies that target virus escape variants. The Journal of Experimental Medicine, 211(12), 2361-2372.
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4

Stereotactic Delivery and Analysis of Transplanted Cells

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We performed stereotactic intracranial injections into the right frontal subcortical white matter (coordinates from bregma: 1.0 mm right – 1.0 mm front – 1.0 mm deep) of NRG (NOD.Cg-Rag1tm1MomIl2rgtm1Wjl/SzJ, Jackson Laboratories) mouse pups at P6–9 as previously described, adapted for mouse pups (Lei et al., 2011 (link)). We injected 12 P5/P6 NRG pups with control and 12 P5/P6 NRG pups with SPARC/SerpinE1-treated cells (~50,000 cells/injection). At 28 and 56 days post-injection, animals were sacrificed and brains were processed for analysis (below). All procedures were performed according to Columbia University IACUC protocol no. AC-AAAV0463.
One month post-transplant, host cortex was sectioned by vibratome at 50 μm. Migratory distance was assessed by scoring tdTomato + cell linear distance from graft site. Two months post-transplant, 300 μm vibratome sections were imaged by confocal microscopy to visualize tdTomato + cell morphology. Processes were traced using ImageJ plugin NeuronJ to assess neurite length and degree of branching.
Genestine M., Ambriz D., Crabtree G.W., Dummer P., Molotkova A., Quintero M., Mela A., Biswas S., Feng H., Zhang C., Canoll P., Hargus G., Agalliu D., Gogos J.A, & Au E. (2021). Vascular-derived SPARC and SerpinE1 regulate interneuron tangential migration and accelerate functional maturation of human stem cell-derived interneurons. eLife, 10, e56063.
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5

Generation of NRG-Flk2−/− and NRG-HLA-A2*0201 Mice

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NRG-Flk2−/− mice were generated by backcrossing mice harboring a Flk2 null allele (Flt3tm1Irl, kindly provided by Ihor Lemishka, Mount Sinai School of Medicine, NY) for 10 generations to NOD Rag1−/−IL2Rgnull mice (NOD.Cg-Rag1tm1MomIl2rgtm1Wjl/SzJ, obtained from the Jackson Laboratory, catalog number 007799). At each generation, the presence of each mutant allele was confirmed with allele-specific primers. Resultant NRG-Flk2+/− were intercrossed to produce NRG-Flk2−/− mice. NRG-HLA-A2*0201 were generated by intercrossing NSG-A2*0201 (NOD.Cg-PrkdcscidIL2rgtmlWjl/Sz Tg(HLA-A2.1)1Eng/Sz)7 (link),9 (link) with NRG mice. The absence of the SCID mutation and the presence of the Rag1, IL2Rγ null alleles and the A2 transgene was determined by PCR. To generated NRG-Flk2−/− HLA-A*0201 (NFA2) mice, NRGF and NRG-A2 mice were intercrossed. NRG, NRG-A2, NRGF, and NFA2 mice were maintained at the Laboratory Animal Resource Center at Princeton University.
All animal experiments described in this study were performed in accordance with protocols (number 1930) that were reviewed and approved by the Institutional Animal Care and Use and Committee of Princeton University.
Douam F., Ziegler C.G., Hrebikova G., Fant B., Leach R., Parsons L., Wang W., Gaska J.M., Winer B.Y., Heller B., Shalek A.K, & Ploss A. (2018). Selective expansion of myeloid and NK cells in humanized mice yields human-like vaccine responses. Nature Communications, 9, 5031.
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6

Xenograft Engraftment in Immunodeficient Mice

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10–14-wk-old NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ), NRG (NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ), or NSGS (NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg[CMV-IL3,CSF2,KITLG]1Eav/MloySzJ; The Jackson Laboratory) mice were irradiated twice with 100 cGy 4 h apart. Mice were injected intraperitoneally with 10% human immunoglobulin solution (1 mg/g of body weight; Privigen; CSL Behring) immediately after the second irradiation. Cells were administered via intratibial injection within 24 h of the second irradiation. Human CD45+33+19− (myeloid) or human CD45+33−19+ (B lymphoid) engraftment was analyzed by FACS and defined as ≥0.1% of live MNC gate. Experiments were performed in accordance with UK government–approved project license 30/2465. Engraftment was monitored by blood or marrow sampling from 12 wk after transplant, and mice were culled for cell harvesting between 12 and 28 wk. Experiments were performed in accordance with UK government–approved project license 30/2465.
Quek L., Otto G.W., Garnett C., Lhermitte L., Karamitros D., Stoilova B., Lau I.J., Doondeea J., Usukhbayar B., Kennedy A., Metzner M., Goardon N., Ivey A., Allen C., Gale R., Davies B., Sternberg A., Killick S., Hunter H., Cahalin P., Price A., Carr A., Griffiths M., Virgo P., Mackinnon S., Grimwade D., Freeman S., Russell N., Craddock C., Mead A., Peniket A., Porcher C, & Vyas P. (2016). Genetically distinct leukemic stem cells in human CD34− acute myeloid leukemia are arrested at a hemopoietic precursor-like stage. The Journal of Experimental Medicine, 213(8), 1513-1535.
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7

Orthotopic Thyroid Tumor Model in Mice

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All animal studies were conducted in accordance with the animal protocol procedures approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Colorado Denver (Aurora, CO). Athymic nude mice (Hsd:Athymic Nude-Foxn1nu (069) from Envigo or in-house breeding for the CUTC60 and CUTC61 cells), SCID/Beige mice (C.B-17/IcrHsd-PrkdcscidLystbg-J from Envigo), or NRG mice (NOD.Cg-Rag1tm1MomIl2rgtm1Wjl/SzJ (#007799) from The Jackson Laboratory) were anesthetized with tribromoethanol (250 mg/kg), and the indicated thyroid cancer cells (5 × 105 in a 5 μL cell suspension) were injected into the right thyroid gland with the aid of a dissecting microscope (Nikon SMZ645), and the skin closed with staples, as previously described (33 (link)–35 (link)). Growth was monitored for the indicated days. Final thyroid tumor size for excised tumors was measured with calipers and volume was calculated using the following formula: tumor volume = (length x width x height)*0.5236.
Schweppe R.E., Pozdeyev N., Pike L.A., Korch C., Zhou Q., Sams S.B., Sharma V., Pugazhenthi U., Raeburn C., Albuja-Cruz M.B., Reigan P., LaBarbera D.V., Landa I., Knauf J.A., Fagin J.A, & Haugen B.R. (2019). Establishment and characterization of four novel thyroid cancer cell lines and PDX models expressing the RET/PTC1 rearrangement, BRAFV600E, or RASQ61R as drivers. Molecular cancer research : MCR, 17(5), 1036-1048.
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8

Establishment of Patient-Derived Xenograft

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Tumor tissue obtained from patient 60 was collected and placed in HBSS (Gibco) on ice for transport. Samples were then cut into pieces approximately 3mm3 with surgical scissors under sterile conditions. Tumor pieces were then transferred to high concentration Matrigel (Corning #354248) and placed on ice until injection. Female NSG mice (NOD.Cg-Rag1tm1MomIl2rgtm1Wjl/SzJ (#007799) from Jackson Laboratory) were anesthetized with 5% inhaled isoflurane in 95% oxygen. Tumors were subcutaneously engrafted into both hind flanks using a 12g trocar needle, and passaged into nude mice at mouse passage 3 (mF3). Tumor growth in vivo was monitored weekly by caliper measurements, with volume calculated using the equation: lengthx(width2)2 .
Schweppe R.E., Pozdeyev N., Pike L.A., Korch C., Zhou Q., Sams S.B., Sharma V., Pugazhenthi U., Raeburn C., Albuja-Cruz M.B., Reigan P., LaBarbera D.V., Landa I., Knauf J.A., Fagin J.A, & Haugen B.R. (2019). Establishment and characterization of four novel thyroid cancer cell lines and PDX models expressing the RET/PTC1 rearrangement, BRAFV600E, or RASQ61R as drivers. Molecular cancer research : MCR, 17(5), 1036-1048.
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9

Human TNBC Mouse Model Development

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A mouse model of human TNBC that has a double mutation in the combining recombinase activating gene (RAG1) and common cytokine receptor γ chain (Cγ) (RAG1xCy) was used as previously described for human pancreatic cancer.3 (link) The RAG1xCγ double mutant mice on a non-obese diabetic (NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ; The Jackson Laboratory) genetic background are completely deficient in T-, B-, and natural killer-cells; show no spontaneous tumor formation; and exhibit normal hematopoietic parameters.15 (link) A breeding colony of mice were generated by intercrossing and were maintained in specific pathogen-free isolators in the Animal Care Facility, Queen’s University, Kingston, ON, Canada. All mice were kept under sterile conditions in microisolators equipped with autoclaved food and water. Experimental protocols using mice in these studies received approval from the Animal Care Committee, Queen’s University. Mice between 6 and 8 weeks of age were used.
Haxho F., Allison S., Alghamdi F., Brodhagen L., Kuta V.E., Abdulkhalek S., Neufeld R.J, & Szewczuk M.R. (2014). Oseltamivir phosphate monotherapy ablates tumor neovascularization, growth, and metastasis in mouse model of human triple-negative breast adenocarcinoma. Breast Cancer : Targets and Therapy, 6, 191-203.
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10

Subcutaneous Tumor Transplantation in Rag1/Il2rg Mice

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We maintained all animals in specific pathogen-free facilities at the Irving Cancer Research Center at CUIMC. All animal procedures are approved by the Institutional Animal Care and Use Committee (IACUC) at CUIMC.
For tumor transplantation assays, we injected 2×106 luciferized HuT78 T-cells subcutaneously into Rag1/Il2rg double knockout recipient mice (NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ, Stock No: 007799, Jackson Laboratory). We monitored tumor development by luciferase bioimaging with the In Vivo Imaging System (IVIS, Xenogen, Alameda, CA).
Cortes J.R., Patrone C.C., Quinn S.A., Gu Y., Sanchez-Martin M., Mackey A., Cooke A., Shih B.B., Laurent A.P., Trager M.H., Ferrando A.A., Geskin L.J, & Palomero T. (2021). JAK-STAT inhibition mediates romidepsin and mechlorethamine synergism in Cutaneous T-cell Lymphoma. The Journal of investigative dermatology, 141(12), 2908-2920.e7.
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