The EFHD1 Knockout AGS Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal population of human AGS gastric epithelial adenocarcinoma cells carrying targeted gene disruption of EFHD1. This knockout model was generated using CRISPR/Cas9-mediated genome editing to introduce loss-of-function mutations in the EFHD1 locus, resulting in a heterogeneous knockout cell pool. The polyclonal format preserves genetic diversity while ensuring functional ablation of EFHD1 protein expression across the population, enabling robust assessment of gene function without clonal artifacts.
The parental AGS cell line was originally derived from a gastric adenocarcinoma and is extensively used as a model system for gastric epithelial biology, Helicobacter pylori pathogenesis, and gastric cancer research. AGS cells exhibit adherent epithelial morphology and retain key signaling pathways relevant to gastric carcinogenesis, including responses to TNF-??, growth factors, and calcium-mobilizing stimuli. Their well-characterized genetic background and suitability for functional assays make AGS an ideal host for studying the role of calcium-binding proteins in gastric cancer.
EFHD1 encodes a calcium-binding protein that localizes to mitochondria and modulates apoptotic signaling through regulation of mitochondrial calcium homeostasis and inter-organellar communication. EFHD1 is activated by TNF-?? and elevated intracellular Ca2+ levels, and it physically interacts with S100A8, S100A9, and F-actin. Downstream, EFHD1 influences the balance of pro-apoptotic BCL2 family members such as BAX and BAK versus anti-apoptotic factors like BCL2. In parallel, EFHD1 modulates NF-??B signaling, impacting transcriptional targets IL-6 and IL-8. The signaling cascade involves calmodulin, CAMKII, NFAT, TNFR1, TRAF2, and I??B??, ultimately regulating cytochrome c release and caspase-3 activation.
In the context of AGS gastric adenocarcinoma cells, disruption of EFHD1 is expected to alter apoptotic sensitivity and calcium-dependent signaling networks. Loss of EFHD1 may compromise mitochondrial calcium buffering, leading to enhanced cytochrome c release and caspase activation upon apoptotic stimuli. Concurrently, EFHD1 knockout could modulate TNF-??-induced NF-??B transcriptional activity, affecting the expression of survival genes and pro-inflammatory cytokines. This makes the model highly relevant for dissecting the interplay between mitochondrial dysfunction and inflammatory signaling in gastric cancer progression and treatment response.
This polyclonal EFHD1 knockout cell product is suited for a broad range of investigational applications, including apoptosis profiling using Annexin V/PI staining and caspase-3/7 activity assays, calcium flux measurements, and NF-??B luciferase reporter gene assays. Researchers can employ transcriptomic approaches such as RNA-seq and quantitative RT-PCR to analyze gene expression changes, while proteomic validation via Western blotting confirms EFHD1 ablation. Functional studies can incorporate MTT viability assays, Transwell migration/invasion tests, and co-immunoprecipitation with S100A8/A9 to probe protein interactions. Additionally, the model supports Helicobacter pylori infection experiments and drug sensitivity screens. For additional information, please contact Ascent Research.
