期刊
VIROLOGY
卷 517, 期 -, 页码 16-23出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.virol.2018.02.019
关键词
Porcine epidemic diarrhea virus (PEDV); Aminopeptidase N (APN); Receptor; CRISPR/Cas9; Vero cells; IPEC-J2 cells
类别
资金
- National Key Research and Development Program of China [2016YFD0500102]
- National Natural Science Foundation of China [31572518]
- Key Research and Development Program of Zhejiang province [2015CO2021]
- Zhejiang Provincial Science Foundation for Distinguished Young Scholars [LR14C180001]
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.
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