Basic Information
-
Gene Targeting Strategy
-
Gene targeting strategy for B-hPVRIG/hTIGIT mice. The exons 1-3 of mouse Pvrig gene that encode the extracellular protein was replaced by human PVRIG exons 2-4 in B-hPVRIG/hTIGIT mice. The exon 2 of mouse Tigit gene that encode the extracellular protein was replaced by human TIGIT exon 2 in B-hPVRIG/hTIGIT mice.
-
mRNA Expression Analysis
-
Strain specific analysis of PVRIG/TIGIT gene expression in wild-type C57BL/6 mice and B-hPVRIG/hTIGIT mice by RT-PCR. Mouse PVRIG/TIGIT mRNA were detectable only in splenocytes of wild-type C57BL/6 mice (+/+). Human PVRIG/TIGIT mRNA were detectable only in homozygous B-hPVRIG/hTIGIT mice (H/H), but not in wild-type C57BL/6 mice.
-
Protein Expression Analysis
-
PVRIG expression analysis in homozygous B-hPVRIG/hTIGIT mice by western blot. Heart was collected from wild type C57BL/6 mice (+/+) and homozygous B-hPVRIG/hTIGIT mice (H/H) and analyzed by western blot with anti-PVRIG antibody. PVRIG was detectable both in wild type C57BL/6 mice and homozygous B-hPVRIG/hTIGIT mice. Anti-PVRIG antibody is crossly reactive with PVRIG in human and mice.
Strain specific TIGIT expression analysis in homozygous B-hPVRIG/hTIGIT mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPVRIG/hTIGIT mice (H/H) stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-TIGIT antibody. Mouse TIGIT was detectable in wild-type mice. Human TIGIT was exclusively detectable in homozygous B-hPVRIG/hTIGIT mice but not in wild-type C57BL/6 mice.
-
Summary
-
mRNA expression analysis:
- Mouse PVRIG/TIGIT mRNA were detectable in splenocytes of wild-type C57BL/6 mice (+/+). Human PVRIG/TIGIT mRNA were detectable in homozygous B-hPVRIG/hTIGIT mice (H/H) but not in wild-type C57BL/6 mice.
Protein expression analysis:
- Mouse PVRIG/TIGIT were detectable in wild-type C57BL/6 mice (+/+). Human PVRIG/TIGIT were detectable in homozygous B-hPVRIG/hTIGIT mice.
