B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice

C57BL/6-Il31tm1(IL31)Bcgen Il31ratm1(IL31RA)BcgenOsmrtm1(OSMR)BcgenTnfrsf4tm1(TNFRSF4)Bcgen Tnfsf4tm1(TNFSF4)Bcgen/Bcgen • 113671

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B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice

Product nameB-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice
Catalog number113671
Strain nameC57BL/6-Il31tm1(IL31)Bcgen Il31ratm1(IL31RA)BcgenOsmrtm1(OSMR)BcgenTnfrsf4tm1(TNFRSF4)Bcgen Tnfsf4tm1(TNFSF4)Bcgen/Bcgen
Strain backgroundC57BL/6
NCBI gene ID386653,133396,9180,7293,7292 (Human)
AliasesIL-31;CRL, CRL3, GLM-R, GLMR, GPL, IL-31RA, PLCA2, PRO21384, hGLM-R, zcytoR17; IL-31R-beta, IL-31RBB, OSMRbeta, PLCA1, OSMR; ACT35, CD134, IMD16, OX40, TXGP1L; CD134L, CD252, GP34, OX-40L, OX4OL, TNLG2B, TXGP1
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  • Description
  • Targeting strategy
  • Phenotypic analysis

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      Description
      • IL-31 signals via a heterodimeric receptor complex composed of a IL‑31 RA and OSMR, and IL-31 directly binds to IL31RA, not OSMR.
      • OX40, also known as CD134 and TNFRSF4, is a 50 kD type I transmembrane glycoprotein.
      • OX40 is a member of the TNF receptor family. OX40 is expressed on activated T lymphocytes including Th1, Th2, Th17, and Treg cells. The interaction of OX40 with OX40L results in B cell proliferation and antibody secretion, regulation of primary T cell expansion, and T cell survival.
      • The mouse Il31 gene that encode the full-length protein was replaced by human IL31 counterpart gene sequences in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • The coding sequences including human IL31RA extracellular region and mouse Il31ra intracellular region were inserted into mouse Il31ra gene in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • The coding sequences including human OSMR extracellular region and mouse Osmr intracellular region were inserted into mouse Osmr gene in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • The exons 1-5 of mouse Ox40 gene that encode extracellular domain are replaced by human counterparts in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • The exons 2-3 of mouse Ox40l gene that  encode the extracellular region were replaced by human OX40L exons 2-3 in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • Mouse OX40 protein was only detectable in wild-type C57BL/6JNifdc mice, and human OX40 protein was only detectable in homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • Mouse OX40L protein was only detectable in wild-type C57BL/6JNifdc mice, and human OX40L protein was only detectable in homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • OSMR protein was detectable in wild-type C57BL/6JNifdc mice and B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • Mouse Il31, Il31ra, Osmr, Ox40 and Ox40l were only detectable in wild-type C57BL/6JNifdc mice. Human IL31, IL31RA, OSMR, OX40 and OX40L mRNA were detectable only in homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • Application: This product is used for pharmacodynamics and safety evaluation of  drugs.
      Targeting strategy

      Gene targeting strategy for B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.

      • The mouse Il31 gene that encode the full-length protein was replaced by human IL31 counterpart gene sequences in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • The coding sequences including human IL31RA extracellular region and mouse Il31ra intracellular region were inserted into mouse Il31ra gene in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • The coding sequences including human OSMR extracellular region and mouse Osmr intracellular region were inserted into mouse Osmr gene in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • The exons 1-5 of mouse Ox40 gene that encode the extracellular domain were replaced by human OX40 exons 1-5 in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      • The exons 2-3 of mouse Ox40l gene that encode the extracellular region were replaced by human OX40L exons 2-3 in B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.
      mRNA expression analysis

      Strain specific analysis of IL31, IL31RA, OSMR, OX40 and OX40L mRNA expression in wild-type C57BL/6JNifdc mice and homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice by RT-PCR. Testis, kidney and spleen RNA were isolated from wild-type C57BL/6JNifdc mice(+/+) and homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice(H/H;H/H;H/H;H/H;H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human IL31, IL31RA, OSMR, OX40 and OX40L primers. Mouse Il31, Il31ra, Osmr, Ox40 and Ox40l mRNA were only detectable in wild-type C57BL/6JNifdc mice. Human IL31, IL31RA, OSMR, OX40 and OX40L mRNA were exclusively detectable in homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice but not in wild-type C57BL/6JNifdc  mice.

      Protein expression analysis-Spleen

      Strain specific OX40 expression analysis in homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc(+/+) and homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice(H/H;H/H;H/H;H/H;H/H) stimulated with anti-CD3ε in vivo, and protein expression was analyzed with anti-mouse OX40 antibody (Biolegend, 119414)  and anti-human OX40 antibody (Biolegend, 350004) by flow cytometry. mOX40 was only detectable in T cells of wild-type C57BL/6JNifdc mice. hOX40 was only detectable in T cells of homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice, but not in wild-type C57BL/6JNifdc mice.

      Protein expression analysis-BMDCs

      Strain specific OX40L expression analysis in homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6JNifdc(+/+) and homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice(H/H;H/H;H/H;H/H;H/H). BMDCs were induced from bone marrow cells and stimulated with LPS. Then BMDCs were analyzed with anti-mouse OX40L antibody (Biolegend, 108814)  and anti-human OX40L antibody (BD Horizon™, 563766) by flow cytometry. mOX40L was only detectable in BMDCs of wild-type C57BL/6JNifdc mice. hOX40L was only detectable in BMDCs of homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice, but not in wild-type C57BL/6JNifdc mice.

      Protein expression analysis

      Strain specific OSMR expression analysis in homozygous humanized B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice by western blot. Brain, lung, kidney, liver, bladder, and ovary were collected from wild-type C57BL/6JNifdc(+/+) and homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice(H/H;H/H;H/H;H/H;H/H) and analyzed by western blot with anti-OSMR antibody(Proteintech, 10982-1-AP). m/hOSMR was detectable in brain, lung, kidney, liver, bladder, and ovary of wild-type C57BL/6JNifdc mice and homozygous B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice.

      * When publishing results obtained using this animal model, please acknowledge the source as follows: The animal model [B-hIL31/hIL31RA/hOSMR/hOX40/hOX40L mice] (Cat# 113671) was purchased from Biocytogen.