The N4BP2L2 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the AGS human gastric adenocarcinoma cell line, designed to disrupt N4BP2L2 gene expression. This product provides a loss-of-function model for studying N4BP2L2 function in an epithelial gastric cancer background. As a polyclonal knockout pool, it maintains the host cell line??s genetic diversity while eliminating target gene expression, enabling robust functional analyses without clonal selection. The knockout is introduced via CRISPR/Cas9-mediated gene disruption, ensuring stable silencing.
The parental AGS cell line, isolated from a human gastric adenocarcinoma, serves as a widely used model in gastric cancer research, including studies of H. pylori infection and inflammatory signaling. AGS cells retain key gastric epithelial characteristics and exhibit robust NF-??B activation upon stimulation, making them an ideal platform for investigating pathways relevant to gastric carcinogenesis.
N4BP2L2 encodes an adaptor protein that recruits NEDD4 family E3 ubiquitin ligases to substrates like TRAF6, promoting ubiquitin-dependent degradation and subsequent downregulation of NF-??B signaling. It interacts with NEDD4, NEDD4L, and ITCH, and operates downstream of inducers such as TNF-??, IL-1??, and cellular stress. By facilitating TRAF6 turnover, N4BP2L2 suppresses NF-??B activation and transcription of target genes including IL-6 and BCL2L1. This positions N4BP2L2 as a negative regulator of the NF-??B pathway, with key components like I??B?? and p65 modulating downstream responses.
In AGS gastric adenocarcinoma cells, disruption of N4BP2L2 is anticipated to enhance NF-??B pathway activity, providing a relevant model for inflammation-associated gastric cancer research. Given the central role of NF-??B in gastric tumorigenesis, this knockout allows investigation of how N4BP2L2 influences cell proliferation, survival, and inflammatory gene expression. The model also offers insights into ulcerative colitis and other inflammation-driven carcinomas where NF-??B signaling is perturbed.
These polyclonal knockout cells are suitable for western blotting of NF-??B phospho-proteins, co-immunoprecipitation of NEDD4 interactions, and ubiquitination assays to study ligase activity. Functional assays such as NF-??B luciferase reporters and RT-qPCR of downstream targets (e.g., IL-6, BCL2L1) can quantify pathway output. Cell proliferation and apoptosis assays further reveal phenotypic consequences of N4BP2L2 loss. The model supports screening for inflammatory signaling modulators and functional genomics of NEDD4 adaptors in gastric cancer. For further technical details or to discuss your specific experimental needs, please contact Ascent Research.
