Strain Name
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C57BL/6N-Crbntm1(CRBN)Bcgen/Bcgen
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Common Name
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B-hCRBN mice
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Background
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C57BL/6N
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Catalog number
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113236
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Aliases
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MRT2, MRT2A
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NCBI Gene ID
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58799
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mRNA expression analysis in humanized B-hCRBN mice
Strain specific analysis of CRBN mRNA expression in wild-type C57BL/6 mice and homozygous B-hCRBN mice by RT-PCR. Heart, liver, spleen, lung, kidney, small intestine, colon and cortex RNA was isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hCRBN mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CRBN primers. Mouse Crbn mRNA was detectable in wild-type C57BL/6 mice. Human CRBN mRNA was detectable only in homozygous B-hCRBN mice but not in wild-type mice.
Protein expression analysis
Strain specific CRBN expression analysis in homozygous B-hCRBN mice by Western blot. The cortex, liver, spleen, lung, kidney were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCRBN mice (H/H), and then analyzed by western blot with anti-CRBN antibody. Mouse CRBN was detectable in wild-type mice. Human CRBN was exclusively detectable in homozygous B-hCRBN mice. The anti-CRBN antibody was cross-reactive between human and mouse.
Functional analysis
Naïve CD4+ T cells from B-hCRBN mice have increased production of IL2 when treated with Lenalidomide, but no change in cells of wild-type mice.
Naïve CD4+ T cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCRBN mice (H/H), then stimulated with DMSO or Lenalidomide in vitro for 24 hours. The supernatants were collected, and IL2 production was measured by ELISA. The results show that Lenalidomide significantly up-regulates the production of IL2 in B-hCRBN mice, but not in wild-type C57BL/6 mice.