• Glycoprotein VI (GPVI) is a 63 kDa platelet/megakaryocyte-specific type I transmembrane glycoprotein of the immunoglobulin superfamily that is an important collagen receptor and initiator of platelet activation, aggregation and thrombin generation.
• The exons 1-8 of mouse Gp6 gene that encode signal peptide, extracellular domain, transmembrane domain and cytoplasmic portion are replaced by human counterparts in B-hGP6 mice. The 3’UTR region of the mouse gene are replaced by human counterparts. The chimeric GP6 expression is driven by endogenous mouse Gp6 promoter, while mouse Gp6 gene transcription and translation will be disrupted.

Gene targeting strategy for B-hGP6 mice. The exons 1-8 of mouse Gp6 gene that encode signal peptide, extracellular domain, transmembrane domain and cytoplasmic portion are replaced by human counterparts in B-hGP6 mice. The 3’UTR region of the mouse gene are replaced by human counterparts. The chimeric GP6 expression is driven by endogenous mouse Gp6 promoter, while mouse Gp6 gene transcription and translation will be disrupted.

Strain specific GP6 expression analysis in homozygous B-hGP6 mice by flow cytometry. Platelets were collected from the blood in wild-type C57BL/6 mice (+/+) and homozygous B-hGP6 mice (H/H). Protein expression was analyzed with anti-human GP6 antibody (BD, 565241) by flow cytometry. Human GP6 was exclusively detectable in homozygous B-hGP6 mice but not in wild-type C57BL/6 mice.

Strain specific analysis of GP6 mRNA expression in wild-type C57BL/6 mice and B-hGP6 mice by RT-PCR. BM RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hGP6 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human GP6 primers. Mouse GP6 mRNA was detectable only in wild-type C57BL/6 mice. Human GP6 mRNA was detectable only in homozygous B-hGP6 mice but not in wild-type mice.