Gene targeting strategy for B-hSEMA7A mice. All exons 1~14 of mouse Sema7a gene that encode the full-length protein were replaced by human SEMA7A exons 1~14 in B-hSEMA7A mice.

Western blot analysis of SEMA7A protein expression in homozygous B-hSEMA7A mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hSEMA7A mice (H/H), and then analyzed by western blot with species-specific anti-SEMA7A antibody. 40 μg total proteins were loaded for western blotting analysis. SEMA7A was detected in brain, testis, lung and colon.

Strain specific analysis of SEMA7A mRNA expression in wild-type C57BL/6 mice and B-hSEMA7A mice by RT-PCR. Brain RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hSEMA7A mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human SEMA7A primers. Mouse Sema7a mRNA was detectable only in wild-type C57BL/6 mice. Human SEMA7A mRNA was detectable only in homozygous B-hSEMA7A mice but not in wild-type mice.