Rapid manipulation of the porcine epidemic diarrhea virus genome by CRISPR/Cas9 technology.
J Virol Methods
; 276: 113772, 2020 02.
Article
in English
| MEDLINE | ID: covidwho-829838
ABSTRACT
Porcine epidemic diarrhea virus (PEDV) is a highly pathogenic enteric coronavirus causing lethal watery diarrhea in suckling piglets. Reverse genetics is a valuable tool to study the functions of viral genes and to generate vaccine candidates. In this study, a full-length infectious cDNA clone of the highly virulent PEDV strain AJ1102 was assembled in a bacterial artificial chromosome (BAC). The rescued virus (rAJ1102) exhibited similar proliferation characteristics in vitro to the wildtype AJ1102. Using CRISPR/Cas9 technology, a recombinant virus rAJ1102-ΔORF3-EGFP in which the ORF3 gene was replaced with an EGFP gene, was successfully generated, and its proliferation characteristics were compared with the parental rAJ1102. Importantly, it just took one week to construct the recombinant PEDV rAJ1102-ΔORF3-EGFP using this method, providing a more efficient platform for PEDV genome manipulation, which could also be applied to other RNA viruses.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Swine Diseases
/
Coronavirus Infections
/
Porcine epidemic diarrhea virus
/
CRISPR-Cas Systems
Topics:
Vaccines
Limits:
Animals
Language:
English
Journal:
J Virol Methods
Year:
2020
Document Type:
Article
Affiliation country:
J.jviromet.2019.113772
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