Basic Information
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Targeting strategy
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Gene targeting strategy for B-hIL31/hIL31RA/hOSMR mice.
The exons 1-3 of mouse Il31 gene that encode the mature protein were replaced by human IL31 exons 1-3 in B-hIL31/hIL31RA/hOSMR mice.
The chimeric IL31RA CDS was inserted in exon 4 of mouse Il31ra gene in B-hIL31/hIL31RA/hOSMR mice.
The coding sequences of human OSMR extracellular region and mouse Osmr intracellular region were inserted into mouse Osmr gene in B-hIL31/hIL31RA/hOSMR mice.
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mRNA expression analysis (IL31/IL31RA)
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Strain specific analysis of IL31 and IL31RA gene expression in wild-type mice and homozygous B-hIL31/hIL31RA mice by RT-PCR. Testis was collect from wild type C57BL/6 mice (+/+) and B-hIL31/hIL31RA mice (H/H). Mouse Il31 and Il31ra mRNA were exclusively detectable in wild-type mice. Human IL31 and IL31RA mRNA were exclusively detectable in homozygous B-hIL31/hIL31RA mice, but not in wild type mice.
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mRNA and protein expression analysis (OSMR)
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Strain specific analysis of OSMR gene expression in wild-type mice and B-hOSMR mice by RT-PCR and western blot. (A) mRNA expression. Mouse Osmr mRNA was exclusively detectable in the kidney of wild-type mice (+/+). Human OSMR mRNA was exclusively detectable in homozygous B-hOSMR mice (H/H). (B) Protein expression. Kidney was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hOSMR mice (H/H), and analyzed by western blot with anti-OSMR antibody. The OSMR protein can be detected in homozygous B-hOSMR mice.
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hIL31 induces pruritus in B-hIL31/hIL31RA/hOSMR mice
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hIL31 induces pruritus in B-hIL31/hIL31RA/hOSMR mice. hIL-31 are administered to homozygous B-hIL31/hIL31RA/hOSMR mice (female, n=3) on Day0, Day3, Day7, Day10 and record scratching counts for 3h after hIL-31 injection. Record scratching counts before hIL-31 injection as baseline. As shown in the panel, hIL-31 induced pruritus successfully in B-hIL31/hIL31RA/hOSMR mice. An antibody targeting IL-31 signaling was used as positive control, which had shown a significant antipruritic effect in our model (Validated by client).
