Description

• Background: CD10 (also known as neutral endopeptidase or NEP) is a zinc-dependent membrane metalloprotease that is widely expressed on the surface of various cells. It plays a key role in renal proteolysis, cardiovascular regulation, and immune responses, as well as in cell proliferation, aging, and fetal development.
• Targeting strategy: In the B-hCD10 rats, exons 3-23 of the rats Cd10 gene, which encode the extracellular domain of CD10, are replaced by the corresponding human CD10 exons.
• Validation: Human CD10 is only detected in homozygous B-hCD10 rats, but not in wild-type SD rats.
• Application: As an early biomarker to predict the onset or progression of early chronic kidney disease (CKD) in patients with type 2 diabetes.

Protein expression analysis

Western blot analysis of CD10 protein expression in homozygous B-hCD10 rats. Various tissue lysates were collected from wild-type SD rats (+/+) and homozygous B-hCD10 rats (H/H), and then analyzed by western blot with cross reactive anti-CD10 antibody (abcam, ab256494). 40 μg total proteins were loaded for western blotting analysis. CD10 was detected in kidneys in both wild-type and homozygous rats.

mRNA expression analysis

Strain specific analysis of CD10 mRNA expression in wild-type SD rats and B-hCD10 rats by RT-PCR. Kidney RNA were isolated from wild-type SD rats (+/+) and homozygous B-hCD10 rats (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with rat or human CD10 primers. Rats Cd10 mRNA were detectable in wild-type SD rats. Human CD10 mRNA was detectable only in homozygous B-hCD10 rats but not in wild-type rats.