LRRC52 is the type I transmembrane protein with an extracellular domain (ECD) containing several LRR motifs and a cytoplasmic C-terminal tail with a short stretch of acidic residues, which is specifically highly expressed in the testis.
The exons 1-2 of mouse Lrrc52 gene that encode signal peptide, and extracellular domain were replaced by human counterparts in B-hLRRC52 mice. The genomic region of mouse Lrrc52 gene that encodes transmembrane domain and cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric LRRC52 expression is driven by endogenous mouse Lrrc52 promoter, while mouse Lrrc52 gene transcription and translation will be disrupted.
LRRC52 protein was detected in testis of homozygous B-h LRRC52 mice.
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mRNA expression analysis in humanized B-hLRRC52 mice
Strain specific analysis of LRRC52 mRNA expression in wild-type C57BL/6 mice and B-hLRRC52 mice by RT-PCR. Testis RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hLRRC52 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human LRRC52 primers. Mouse Lrrc52 mRNA was detectable only in wild-type C57BL/6 mice. Human LRR52 mRNA was detectable only in homozygous B-hLRRC52 mice but not in wild-type mice.
Protein expression analysis
Strain specific LRRC52 expression analysis in homozygous B-hLRRC52 mice by western blot. Heart,lung, kidney, testis, and brain were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hLRRC52 mice (H/H), and analyzed by western blot with anti-LRRC52 antibody(Invitrogen,PA5-107159). m/hLRRC52 was detectable in testis of wild-type C57BL/6 mice (+/+) and homozygous B-hLRRC52 mice.