Tollip敲除猪肾细胞系的构建

doi:10.11843/j.issn.0366-6964.2024.04.042

Abstract

Abstract: In order to explore the effect of Toll-interacting protein (Tollip) on the proliferation of Foot-and-mouth disease virus (FMDV), CRISPR/Cas9 gene editing technology was used to construct a porcine kidney cell line (PK-15) with Tollip gene deletion, and the knockout effect was confirmed by PCR sequencing and Western blot, Then, CCK-8 kit was used to determine the change of cell viability after Tollip deletion, and there was no obvious difference between knockout cells and wild cells. Western blot, indirect immunofluorescence, real-time fluorescence and viral titer were used to determine the replication of viruses in knockout cell lines during FMDV infection. The results showed that knockout cells significantly promoted virus replication compared with wild cells during FMDV infection. In summary, Tollip deletion cell lines were successfully constructed on PK-15 cells, and the results showed that Tollip deletion contributed to FMDV replication. These results provide a theoretical basis for the subsequent study of Tollip function and antiviral mechanism.

Key words: CRISPR/Cas9, Tollip gene, cell lines, foot-and-mouth disease virus

CLC Number:

Q789

WANG Jiali, YANG Fan, SHAO Wenhua, HUANG Mengyao, CAO Weijun, PU Xiuying, ZHANG Wei, ZHENG Haixue. Construction of Tollip Knockout Pig Kidney Cell Line[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1810-1818.

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Construction of Tollip Knockout Pig Kidney Cell Line

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