Targeting strategy
Gene targeting strategy for B-hGPR55 mice. The exon 2 of mouse Gpr55 gene that encodes the full-length protein was replaced by human GPR55 exon 2 in B-hGPR55 mice.
mRNA expression analysis
Strain specific analysis of GPR55 mRNA expression in wild-type C57BL/6 mice and B-hGPR55 mice by RT-PCR. Brain RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hGPR55 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human GPR55 primers. Mouse Gpr55 mRNA was detectable only in wild-type C57BL/6 mice. Human GPR55 mRNA was detectable only in homozygous B-hGPR55 mice but not in wild-type mice.
Protein expression analysis
Western blot analysis of GPR55 protein expression in homozygous B-hGPR55 mice. Various tissue lysates were collected from C57BL/6N wildtype (+/+) mice and homozygous B-hGPR55 (H/H) mice, and then analyzed by western blot with anti-GPR55 antibody. 40 μg total proteins were loaded for western blotting analysis. GPR55 was detected in brain, pancreas, liver and stomach in homozygous B-hGPR55 mice. （The arrow above indicates the glycosylated GPR55 protein, the arrows below indicates the GPR55 protein）
* When publishing results obtained using this animal model, please acknowledge the source as follows: The animal model [B-hGPR55 mice] (Cat# 112462) was purchased from Biocytogen.
