Establishment of a STING‐Deficient HepG2 Cell Line through CRISPR/Cas9 System and Evaluation of Its Effects on Salmonella Replication

Author:

Sun LanqingORCID,Huang KaiORCID,Huang Xuan

Abstract

Background. Salmonella enterica serovar Typhimurium (Salmonella Typhimurium) is a common food‐borne pathogen that causes gastroenteritis and can lead to life‐threatening systemic disease when it spreads to vital organs, such as the liver. Stimulator of interferon genes (STING) is a crucial regulator of the host’s innate immune response to viral infections, while its role in bacterial infections remains controversial. This study aims to establish a STING‐deficient HepG2 cell line through the CRISPR/Cas9 system and evaluate its effects on Salmonella replication. Methods. In this study, a STING knockout HepG2 cell line was constructed through the application of CRISPR/Cas9 technology. We assessed cell viability and proliferation using the CCK‐8 assay. Subsequently, we investigated the effect of STING deletion on Salmonella replication and the expression of type I interferon‐related genes. Results. The STING knockout HepG2 cell line was successfully constructed using the CRISPR/Cas9 system. The proliferation capability was diminished in STING‐deficient HepG2 cells, while Salmonella Typhimurium replication in these cells was augmented compared to the wild‐type (WT) group. Following Salmonella infection, the transcriptional responses of type I interferon‐related genes, such as IFNB1 and ISG15, were inhibited in STING‐deficient HepG2 cells. Conclusions. We successfully constructed a STING‐deficient cell line. Our finding of increased Salmonella Typhimurium replication in STING‐deficient HepG2 cells provides the basis for further studies on pathogen‐host interactions.

Publisher

Wiley

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