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Construction of cell lines with ring finger protein 31 gene knockout and evaluation of its impact on the replication of foot-and-mouth disease virus
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Tingting ZHOU1, 2, Wei ZHANG2, Huanan LIU2, Zhitong CHEN2, Xingyan DONG2, Songhao WANG3, Weijun CAO2, Haixue ZHENG1, 2, Xiuying PU1, Fan YANG2
Acta Microbiologica Sinica | 2026, 66(5) : 2384 - 2392
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Acta Microbiologica Sinica | 2026, 66(5): 2384-2392
• Research Article •
Construction of cell lines with ring finger protein 31 gene knockout and evaluation of its impact on the replication of foot-and-mouth disease virus
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Tingting ZHOU1, 2, Wei ZHANG2, Huanan LIU2, Zhitong CHEN2, Xingyan DONG2, Songhao WANG3, Weijun CAO2, Haixue ZHENG1, 2, Xiuying PU1, Fan YANG2
Affiliations
  • 1.School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, Gansu, China
  • 2.State Key Laboratory of Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China
  • 3.Jiaxian Bureau of Agriculture and Rural Affairs, Pingdingshan, Henan, China
Published: 2026-05-04 doi: 10.13343/j.cnki.wsxb.20250874
Outline
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Objective To investigate the function of ring finger protein 31 (RNF31) in the replication of foot-and-mouth disease virus (FMDV) and to provide a theoretical basis for the research on the molecular mechanism by which the host protein RNF31 regulates FMDV replication. Methods CRISPR/Cas9 gene editing was employed to design two sgRNA sequences in the exon segment of RNF31, and recombinant plasmids were constructed by ligation with the pX459-puro vector. The recombinant plasmids pX459-RNF31-sgRNA were transfected into PK-15 cells, followed by screening under the action of puromycin to obtain the cell lines with RNF31 gene knockout. The effect of RNF31 gene knockout on FMDV replication was detected by Western blotting, RT-qPCR, and TCID50 methods. Results Compared with wild-type cells, the knockout of RNF31 significantly increased the protein level, mRNA level, and virus titer of FMDV. Conclusion We successfully construct the cell lines with RNF31 gene knockout and prove that RNF31 plays a key role in the replication of FMDV. This result provides data support for further research on the mechanism by which RNF31 inhibits FMDV replication.

CRISPR/Cas9  /  ring finger protein 31  /  knockout cell lines  /  foot-and-mouth disease virus
Tingting ZHOU, Wei ZHANG, Huanan LIU, Zhitong CHEN, Xingyan DONG, Songhao WANG, Weijun CAO, Haixue ZHENG, Xiuying PU, Fan YANG. Construction of cell lines with ring finger protein 31 gene knockout and evaluation of its impact on the replication of foot-and-mouth disease virus[J]. Acta Microbiologica Sinica, 2026 , 66 (5) : 2384 -2392 . DOI: 10.13343/j.cnki.wsxb.20250874
Funding
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  • The National Natural Sciences Foundation of China(32330107)
  • The Gansu Provincial Joint Research Fund(25JRRA1088)
  • The Gansu Provincial Joint Research Fund(25JRRA1087)
  • The Gansu Province Research and Development-Industry Integration and Technology Empowerment Program(25FNNA002)
  • The Gansu Provincial Major Science and Technology Special Program(22ZD6NA001)
  • The Earmarked Fund for CARS-35, and the Project of National Center of Technology Innovation for Pigs(CARS-35)
  • The Earmarked Fund for CARS-35, and the Project of National Center of Technology Innovation for Pigs(NCTIP-XD/C03)
Appendix
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Year 2026 volume 66 Issue 5
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110
48
Cite this Article
BibTeX
Article Info
doi: 10.13343/j.cnki.wsxb.20250874
  • Receive Date：2025-11-23
  • Online Date：2026-05-09
  • Published：2026-05-04
Article Data
Affiliations
History
  • Received：2025-11-23
  • Accepted：2026-01-19
Funding
The National Natural Sciences Foundation of China(32330107)
The Gansu Provincial Joint Research Fund(25JRRA1088)
The Gansu Provincial Joint Research Fund(25JRRA1087)
The Gansu Province Research and Development-Industry Integration and Technology Empowerment Program(25FNNA002)
The Gansu Provincial Major Science and Technology Special Program(22ZD6NA001)
The Earmarked Fund for CARS-35, and the Project of National Center of Technology Innovation for Pigs(CARS-35)
The Earmarked Fund for CARS-35, and the Project of National Center of Technology Innovation for Pigs(NCTIP-XD/C03)
Affiliations
    1.School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, Gansu, China
    2.State Key Laboratory of Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China
    3.Jiaxian Bureau of Agriculture and Rural Affairs, Pingdingshan, Henan, China

Corresponding:

E-mail: PU Xiuying, ;
YANG Fan,
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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