Mice were housed in AAALAC-accredited vivarium. Room temperature was maintained at 22 ± 3 °C and the humidity at 55 ± 15%. Animals were kept at a 12 h/12 h dark/light cycle. The C57BL/6 and SCID/Beige mice were purchased from Charles River (Wilmington, MA). Nude mice, Col1a2-CreERT2, C57BL/6-Tg(TRAMP)8247Ng/J, and B6.Cg-Gt(ROSA)26Sortm3(CAG-EYFP)Hze/J, Ptentm1Hwu, and R26-LSL-KrasG12D mice were purchased from the Jackson Laboratory (Bar Harbor, ME). The ARR2Pb-Cre mice were from Dr. Fen Wang at the Institute of Biosciences and Technology at Texas A&M. The R26-LSL-Foxf2 mice were generated at the Baylor College of Medicine. The Foxf2 null mouse embryos were generously provided by Dr. Rulang Jia at Cincinnati Children’s Hospital. All mice were on the C57BL/6 background. Mice were genotyped by polymerase chain reaction using mouse genomic DNA from tail biopsy specimens. The sequences of genotyping primers and the expected band sizes for PCR are listed in Supplementary Table 2 . PCR products were separated electophoretically on 1% agarose gels and visualized via ethidium bromide under UV light.
Col1a2 creert2
Manufactured by Jackson ImmunoResearch
Sourced in United States
Col1a2-CreERT2 is a Cre-recombinase transgene that is expressed under the control of the Col1a2 promoter. The Cre-recombinase is fused to a mutated estrogen receptor (ERT2) domain, which provides ligand-dependent Cre activity.
Automatically generated - may contain errors
Lab products found in correlation
Scid beige, by Charles River Laboratories (5 mentions)
B6 cg gt rosa 26sortm3 cag eyfp hze j, by Jackson ImmunoResearch (5 mentions)
C57bl 6, by Charles River Laboratories (5 mentions)
C57bl 6 tg tramp 8247ng j, by Jackson ImmunoResearch (2 mentions)
Pdgfrβ creert2, by Jackson ImmunoResearch (2 mentions)
Lgr5 dtr egfp mice, by Genentech (2 mentions)
B6.129p2 lgr5tm1 cre ert2 cle j, by Jackson ImmunoResearch (2 mentions)
Ng2 creert2, by Jackson ImmunoResearch (2 mentions)
C57bl 6 gt rosa 26sortm1 hbegf awai j, by Jackson ImmunoResearch (2 mentions)
B6 cg gt rosa 26sortm14 cag tdtomato hze j, by Jackson ImmunoResearch (2 mentions)
Ifnar1fl fl, by Jackson ImmunoResearch (2 mentions)
Tamoxifen, by Merck Group (2 mentions)
Ptentm1hwu, by Jackson ImmunoResearch (1 mentions)
129s wlstm1.1lan j, by Jackson ImmunoResearch (1 mentions)
Acta2 creert2, by Jackson ImmunoResearch (1 mentions)
Rosa26 lsl tdtomato, by Jackson ImmunoResearch (1 mentions)
Hif1α floxed mice, by Jackson ImmunoResearch (1 mentions)
C57bl 6 mice, by Charles River Laboratories (1 mentions)
11 protocols using col1a2 creert2
1
Genotyping of Transgenic Mouse Lines
2
Conditional Knockout of IL-33 in Mice
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Male mice carrying a floxed IL‐33 allele (IL‐33fl/fl; Jackson Laboratory, Bar Harbor) were bred with homozygote mice carrying transgenes for COL1A2 Cre‐ERT2 (Jackson Laboratory) or Aggrecan Cre‐ERT2 (Jackson Laboratory). Hence, we generated two different types of homozygote double transgenic tissue‐specific conditional knockout (KO) mice. (1) IL‐33fl/fl; COL1A2 Cre‐ERT2 (IL‐33COL1A2 Cre‐ERT2) that are synovial‐specific IL‐33‐conditional KO mice. (2) Aggrecan Cre‐ERT2 (IL‐33Acan Cre‐ERT2) that are cartilage‐specific IL‐33‐conditional KO mice. Mice were obtained at the expected Mendelian ratio with no adverse phenotypic side effects. Adult mice (8 weeks of age) were administered intraperitoneal doses of free base tamoxifen (TX) (2 mg kg−1; Sigma‐Aldrich) three times every other day for one week to induce conditional KO of IL‐33. IL‐33fl/fl mice were used as controls for conditional KO mice unless otherwise stated.
3
Genotyping Transgenic Mouse Models
4
Lineage Tracing and Gene Ablation in Prrx1-CreERT2 Mice
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The knock-in Prrx1CreERT2 mouse line was generated by Shanghai Biomodel Organism Science & Technology Development Co., Ltd. In this study, heterozygous Prrx1CreERT2 mice were used for lineage tracing or gene ablation. Rosa26LSL-tdTomato, Acta2-CreERT2, and Col1a2-CreERT2 mice were purchased from the Jackson Laboratory (https://www.jax.org/cn/ ). Bmpr1af/f mice were generated in Yuji Mishina’s laboratory. Animal experiments were carried out in accordance with recommendations in the National Research Council Guide for Care and Use of Laboratory Animals and in comply with relevant ethical regulations for animal testing and research, with the protocols approved by the Institutional Animal Care and Use Committee of Shanghai, China (SYXK(SH)2011–0112). This study follows the project that ‘Studies on the mechanism of MSC self-renewal differentiation and regulation of related tissue stem cells,’ which was approved by Shanghai Jiao Tong University in 2015 and 2024, the approval number is A2015027 and A2024014. Male mice were used in this study. Mice were anesthetized using 40 mg kg–1 sodium pentobarbital by intraperitoneal injection or euthanized by carbon dioxide inhalation.
5
Genetically Engineered Mouse Models
6
Conditional Knockout Mouse Model for Hypoxia Signaling
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All experiments were conducted in accordance with Institutional Animal Care and Use Committee guidelines at the Cleveland Clinic Lerner Research Institute. The Col1a2-CreERT2 (Zheng et al., 2002 (link)) and HIF1α floxed mice (Ryan et al., 2000 (link)) were purchased from The Jackson Laboratory. IL-17RC floxed mice were generated by Cyagen Biosciences, with loxP sites flanking exons 6–7 of Il17rc. Gender- and age-matched mice were used for all experiments.
To induce transient Cre activity, tumor-bearing mice were i.p. injected with TAM (∼5 mg/25 g weight for single dose or ∼1 mg/25 g weight, three doses as indicated, e.g., days 0, 7, and 14 of the syngeneic model) for each experiment.
To induce transient Cre activity, tumor-bearing mice were i.p. injected with TAM (∼5 mg/25 g weight for single dose or ∼1 mg/25 g weight, three doses as indicated, e.g., days 0, 7, and 14 of the syngeneic model) for each experiment.
7
Genetically Engineered Mouse Models
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All animals used in this study received humane care in compliance with the principles stated in the Guide for the Care and Use of Laboratory Animals, NIH Publication, 1996 edition, and the protocol was approved by the Institutional Animal Care Committee of Universityof Washington. The C57BL/6 and SCID/Beige mice were purchased from Charles River (Wilmington, MA). Lgr5-DTR-eGFP mice were originally generated by Genentech (Tian et al., 2011 (link)) and were kindly provided by Dr. Noah Shroyer at the Baylor College of Medicine. Krt7-CreERT2 was kindly provided by Dr. Jianwen Que at Columbia University. Krt8-CreERT2 was made by our laboratory as described (Zhang et al., 2012 ). C57BL/6-Gt(ROSA)26Sortm1(HBEGF)Awai/J, B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J, B6.129P2-Lgr5tm1(cre/ERT2)Cle/J, Col1a2-CreERT2, Pdgfrβ-CreERT2, NG2-CreERT2, B6.Cg-Gt(ROSA)26Sortm3(CAG-EYFP)Hze/J mice were from the Jackson Laboratory (Bar Harbor, ME). Male mice at the age of E15.5 to postnatal 27 weeks were used. Mice were genotyped by polymerase chain reaction using mouse genomic DNA from tail biopsy specimens. The sequences of genotyping primers and the expected band sizes for PCR are listed in Table S3 . PCR products were separated electrophoretically on 1% agarose gels and visualized via ethidium bromide under UV light.
8
Conditional Knockout of IFNAR1 in Mice
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All mouse experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of The University of Pennsylvania. Mice were maintained in a specific pathogen-free facility in accordance with the American Association for Laboratory Animal Science guidelines. All mice were of C57BL/6NJ background, and unless otherwise described are 6-8 weeks of age. Mice of both sexes were utilized for these studies. Rag1−/−, Ifnar1fl/fl, and Col1a2-CreERT2/+ mice were obtained from Jackson Laboratory (Bar Harbor, ME, USA). Ifnar1S526A mice (“SA”) and bone marrow chimeras were generated as previously described [23 (link)]. Fibroblast-specific IFNAR1 knockout mice (Ifnar1Δfib) were established by crossing Ifnar1fl/fl mice with Col1a2-CreERT2/+ mice. The resultant Col1a2-CreERT2/+; Ifnar1fl/fl mice were genotyped by PCR (Figure 3I ). The Col1a2-CreERT2/+ was induced by administering tamoxifen (Sigma #T5648, St. Louis, MO, USA) 0.2 mg/g, daily for 5 days. Deletion of IFNAR1 was confirmed by PCR with primers for mouse wildtype IFNAR1 (Forward: 5’-CTGGGAGCCAGGGCATAAC-3’, Reverse 5’-CCAGCCTTTCGGAGTGTGC-3’) (Figure 3I ). Littermates were randomly assigned into experimental groups, which were either co-housed or exposed to other groups’ bedding to ensure exposure to all groups’ microbiota.
9
Conditional Knockout of IFNAR1 in Mice
10
Genetically Modified Mouse Models
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