The CD38 Knockout AGS Polyclonal Cells represent a pool of CRISPR/Cas9-edited AGS cells carrying targeted disruptions in the CD38 gene, resulting in a heterogeneous population of CD38-null gastric epithelial cells. This polyclonal knockout format preserves genetic diversity while ensuring functional ablation of CD38, providing a versatile loss-of-function model for interrogating CD38-dependent molecular mechanisms without the clonal artifacts associated with monoclonal isolates.
The AGS cell line, isolated from a human gastric adenocarcinoma, is an adherent epithelial line extensively utilized to model gastric cancer biology, including studies of Helicobacter pylori infection, oncogenic signaling, and epithelial-mesenchymal transition. AGS cells retain key signaling pathways relevant to gastric tumorigenesis, making them an appropriate host for dissecting cancer-relevant functions of CD38.
CD38 (cluster of differentiation 38) is a type II transmembrane glycoprotein that functions as a bifunctional ectoenzyme with ADP-ribosyl cyclase and NAD+ glycohydrolase activities, catalyzing the conversion of NAD+ to cyclic ADP-ribose (cADPR) and ADP-ribose. cADPR serves as a second messenger that mobilizes calcium from intracellular stores by activating ryanodine receptors (RyR) and, indirectly, IP3 receptors. Beyond its enzymatic role, CD38 acts as a receptor mediating cell adhesion through interactions with CD31 (PECAM-1) and associating with caveolin-1 and integrins. In AGS cells, CD38 signaling is activated by inflammatory cytokines such as TNF-?? and IFN-??, as well as by Helicobacter pylori infection, and converges on downstream effectors including calcium/calmodulin-dependent kinase II (CaMKII), Akt, ERK1/2, and NF-??B, collectively promoting proliferative and survival signals.
Disruption of CD38 in AGS cells abrogates CD38-dependent NAD+ catabolism, leading to decreased intracellular cADPR levels and attenuated calcium mobilization. This dampens downstream PI3K/Akt and NF-??B signaling cascades, which are critical drivers of gastric cancer cell proliferation, survival, and migration. Consequently, the CD38 knockout AGS model enables the direct interrogation of CD38??s contributions to gastric adenocarcinoma pathobiology, including its role in maintaining malignant phenotypes and mediating responses to microenvironmental cues.
Researchers can employ this polyclonal knockout population in a wide array of experimental contexts, including calcium imaging to monitor cADPR-mediated calcium fluxes, phospho-specific western blotting to assess Akt and ERK activation, and functional assays such as MTT proliferation, transwell migration, and Annexin V apoptosis. Additional applications encompass co-immunoprecipitation of CD31 and caveolin-1 complexes, NAD+ quantification, and transcriptomic profiling via RNA-seq. This model is a valuable tool for studies on gastric cancer signaling, drug resistance, and H. pylori pathogenesis. For further inquiries, contact Ascent Research.
