Gene targeting strategy for B-hPSMA mice. The extracellular coding sequence of human PSMA gene and mouse 3’ UTR were inserted into mouse Psma exon 2 in B-hPSMA mice, so the mouse Psma gene was disrupted. The intracellular and transmembrane sequence of mouse Psma did not change.
Protein expression analysis
IHC for human PSMA expression. Kidney tissues from wild-type (+/+) and B-hPSMA mice (H/H) were stained with an antibody specific for human PSMA, and only the kidney tubular epithelial cells from B-hPSMA mice (H/H) showed hPSMA expression. (C57BL/6-1: kidney tissues from C57BL/6 mice were only stained with anti-IgG antibody; Human,C57BL/6-2 and B-hPSMA mice kidney tissues were stained with anti-human PSMA antibody.)
mRNA expression analysis (data not shown, please check the data in technical confidential version of PPT ):
1. Human PSMA was detectable in B-hPSMA mice (H/H), and confirms that the transcript is complete by sequencing.
2. Mouse Psma still exsits, but it’s not sure if it can express the normal protein for many transcripts. Considering our targeting strategy, the mouse Psma gene had been genetically damaged, so it should not express the protein.
Protein expression analysis:
1.Human PSMA was detectable on the kidney tubular epithelial cells of B-hPSMA mice (H/H).
2. There were no antibodies that specifically recognize mouse Psma, so we didn’t detect the mouse PSMA protein expression in B-hPSMA mice (H/H).