Aminopeptidase-N-independent entry of porcine epidemic diarrhea virus into Vero or porcine small intestine epithelial cells.
A monkey cell line Vero (ATCC CCL-81) is commonly used for porcine epidemic diarrhea virus (PEDV) propagation in vitro. However, it is still controversial whether the porcine aminopeptidase N (pAPN) counterpart on Vero cells (Vero-APN) confers PEDV entry. We found that endogenous expression of Vero-APN was undetectable in the mRNA and the protein levels in Vero cells. We cloned the partial Vero-APN gene (3340-bp) containing exons 1 to 9 from cellular DNA and subsequently generated two APN-knockout Vero cell lines by CRISPR/Cas9 approach. PEDV infection of two APN-knockout Vero cells had the same efficiency as the Vero cells with or without neuraminidase treatment. A Vero cells stably expressing pAPN did not increase PEDV production. SiRNA-knockdown of pAPN in porcine jejunum epithelial cells had no effects on PEDV infection. The results suggest that there exists an additional cellular receptor on Vero or porcine jejunal cells independent of APN for PEDV entry.
['Ji|Chun-Miao|CM|', 'Wang|Bin|B|', 'Zhou|Jiyong|J|', 'Huang|Yao-Wei|YW|']
D000818:Animals D000914:Antibodies, Viral D018826:CD13 Antigens D002522:Chlorocebus aethiops D004847:Epithelial Cells D017353:Gene Deletion D007421:Intestine, Small D053485:Porcine epidemic diarrhea virus D064370:Spike Glycoprotein, Coronavirus D013552:Swine D014709:Vero Cells D053586:Virus Internalization
Porcine epidemic diarrhea virus (PEDV) Vero cells CRISPR/Cas9 Receptor Aminopeptidase N (APN) IPEC-J2 cells