Basic Information
-
Targeting strategy
-
Gene targeting strategy for B-hCMKLR1 mice.
The exon 3 of mouse Cmklr1 gene that encodes the full-length protein was replaced by human CMKLR1 exons 3-4 in B-hCMKLR1 mice.
-
mRNA expression analysis
-
Strain specific analysis of CMKLR1 mRNA expression in wild-type C57BL/6 mice and B-hCMKLR1 mice by RT-PCR. Spleen RNA was isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hCMKLR1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CMKLR1 primers. Mouse Cmklr1 mRNA was detectable in wild-type C57BL/6 mice. Human CMKLR1 mRNA was detectable only in homozygous B-hCMKLR1 mice but not in wild-type mice. Specially, the human CMKLR1 isoform b was actually transcribed in B-hCMKLR1 mice.
-
Protein expression analysis
-
Strain specific CMKLR1 expression analysis in homozygous B-hCMKLR1 mice by flow cytometry. Splenocytes, blood, lymph modes (LN) and bone marrow (BM) were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCMKLR1 mice (H/H), and analyzed by flow cytometry with species-specific anti-CMKLR1 antibody. mCMKLR1 was detectable in wild-type mice, hCMKLR1 was detectable in homozygous B-hCMKLR1 mice.
Strain specific CMKLR1 expression analysis in homozygous B-hCMKLR1 mice by flow cytometry. Splenocytes, blood and bone marrow (BM) were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCMKLR1 mice (H/H), and analyzed by flow cytometry with species-specific anti-CMKLR1 antibody. mCMKLR1 was detectable in wild-type mice, hCMKLR1 was detectable in homozygous B-hCMKLR1 mice.
