Basic Information
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Description
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- IL1R1 encodes a cytokine receptor that belongs to the interleukin-1 receptor family. The encoded protein is a receptor for interleukin-1 alpha, interleukin-1 beta, and interleukin-1 receptor antagonist. It is an important mediator involved in many cytokine-induced immune and inflammatory responses. IL1R1 plays a key role in the immune system and is an important research target in tumors, autoimmune diseases, and central nervous system disorders.
- Gene targeting strategy for B-hIL1R1 mice. A chimeric CDS that encodes human IL1R1 signal peptide and extracellular domain, mouse Il1r1 transmembrane and cytoplasmic domain is inserted right after mouse Il1r1 exon 2 to replace the part of exon 2 to exon 5 of mouse Il1r1 gene. The chimeric IL1R1 protein expression will be driven by endogenous mouse Il1r1 promoter, while mouse Il1r1 gene transcription and translation will be disrupted.
- Human IL1R1 mRNA was detectable only in homozygous B-hIL1R1 mice. Human IL1R1 was exclusively detectable in homozygous B-hIL1R1 mice. And IL1B can trigger downstream signaling pathways in B-hIL1R1 mice.
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Targeting strategy
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Gene targeting strategy for B-hIL1R1 mice. A chimeric CDS that encodes human IL1R1 signal peptide and extracellular domain, mouse Il1r1 transmembrane and cytoplasmic domain is inserted right after mouse Il1r1 exon 2 to replace the part of exon 2 to exon 5 of mouse Il1r1 gene. The chimeric IL1R1 protein expression will be driven by endogenous mouse Il1r1 promoter, while mouse Il1r1 gene transcription and translation will be disrupted.
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mRNA expression analysis in humanized B-hIL1R1 mice
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Species specific analysis of IL1R1 gene expression in wild-type C57BL/6 mice and homozygous humanized B-hIL1R1 mice by RT-PCR. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL1R1 mice (H/H). Mouse Il1r1 mRNA was detectable only in wild-type C57BL/6 mice. Human IL1R1 mRNA was detectable only in homozygous B-hIL1R1 mice, but not in wild-type C57BL/6 mice.
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Protein expression analysis in bone marrow
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Strain specific IL1R1 expression analysis in wild-type C57BL/6 mice and homozygous B-hIL1R1 mice by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL1R1 mice (H/H), and cultured in 6-well plates stimulated with GM-CSF and IL-4 for 7 days to induce BMDCs. Then BMDCs were collected and analyzed by flow cytometry with anti-hIL1R1 antibody (R&D, FAB269A-25). Human IL1R1 was exclusively detectable in homozygous B-hIL1R1 mice.
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Functional analysis
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Function analysis of IL1R1 in wild-type C57BL/6 mice and homozygous B-hIL1R1 mice by ELISA. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL1R1 mice (H/H), and then stimulated with 100 ng/mL mouse IL1B (mIL1B) or human IL1B (hIL1B) for 48 h. Both mIL1B and hIL1B can induce mouse IL-6 production in wild-type C57BL/6 mice and homozygous B-hIL1R1 mice. The results indicate that IL1B can trigger downstream signaling pathways in B-hIL1R1 mice.
