Interference effect of CRISPR/Cas13b on porcine epidemic diarrhea virus

MEI Yan; YANG Huaqiang; WU Zhenfang

doi:10.7671/j.issn.1001-411X.201811022

Journal of South China Agricultural University > 2019 > 40(6): 1-7. > DOI: 10.7671/j.issn.1001-411X.201811022

MEI Yan, YANG Huaqiang, WU Zhenfang. Interference effect of CRISPR/Cas13b on porcine epidemic diarrhea virus[J]. Journal of South China Agricultural University, 2019, 40(6): 1-7. DOI: 10.7671/j.issn.1001-411X.201811022

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Interference effect of CRISPR/Cas13b on porcine epidemic diarrhea virus

MEI Yan^1,,
YANG Huaqiang^{1, 2,},
WU Zhenfang^{1, 2, ,}

1.
College of Animal Science, South China Agricultural University, Guangzhou 510642, China
2.
Wens’ Institute, Yunfu 527400, China

More Information

Received Date: November 13, 2018
Accepted Date: October 27, 2019
Available Online: May 17, 2023

Graphical Abstract

Abstract

Abstract

Objective
Porcine epidemic diarrhea, caused by porcine epidemic diarrhea virus (PEDV), is a highly contagious viral disease and results in high mortality of pigs and huge lost of pig industry. The CRISPR/Cas13b system can mediate a highly efficient cleavage or editing to target RNA, thereby offering a novel strategy for interfering the infection of RNA viruses. We here tried to use the CRISPR/Cas13b system to cleave the PEDV RNA genome, in order to explore a novel strategy to inhibit PEDV infection.

Method
We designed four CRISPR RNA (crRNA) sequences which respectively recognize four regions in the PEDV genome. The CRISPR/Cas13b targeting vectors were constructed and transfected into Vero cells. The transfected cells were infected by PEDV, and then we analyzed the viral load of PEDV in cultured cells.

Result
The CRISPR/Cas13b system significantly inhibited PEDV propagation in Vero cells. In the viral immunofluorescence assay, the transfected cells with targeting vectors U6-crRNA3 and U6-crRNA4 had obviously fewer fluorophores compared with normal cells. The quantitative PCR results showed that CRISPR/Cas13b decreased PEDV load in cultured cells by above 50%.

Conclusion
The constructed CRISPR/Cas13b system can effectively interfere the propagation of PEDV. This study provides an alternative approach for effective RNA virus prevention and control, and creation of disease-resistant pig models.
- pig,
- porcine epidemic diarrhea virus,
- CRISPR/Cas13b system,
- targeting plasmid,
- RNA virus,
- gene editing

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References (22)

References

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Interference effect of CRISPR/Cas13b on porcine epidemic diarrhea virus