The ELANE knockout AGS polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS gastric adenocarcinoma cell line, engineered for loss-of-function studies of the ELANE gene. This polyclonal pool offers a genetically heterogeneous background that minimizes clonal artifacts while enabling robust functional interrogation of ELANE. The use of CRISPR/Cas9-mediated gene disruption generates a versatile knockout model suitable for a range of cell-based assays.
AGS is a widely used human cell line established from a gastric adenocarcinoma, providing a well-characterized epithelial model of gastric cancer biology. These cells retain key features of gastric epithelium and are extensively employed to investigate gastric carcinogenesis, host-pathogen interactions, and mucosal inflammation. The AGS background supports studies of signaling pathways relevant to gastric cancer progression and inflammatory responses.
ELANE encodes neutrophil elastase, a serine protease primarily stored in azurophil granules of neutrophils and released upon activation. This enzyme degrades extracellular matrix components such as elastin and collagens, cleaves bacterial proteins, and modulates inflammatory signaling by proteolytically processing cytokines and receptors. ELANE activity is regulated by upstream factors including G-CSF (CSF3) and inflammatory cytokines (TNF, IL-1??), and is transcriptionally controlled by C/EBP?? and PU.1. Downstream targets comprise IL-8, TNF, MMP9, and CD14, with receptor crosstalk via PAR-2. Key interacting partners include the endogenous inhibitors ??1-antitrypsin (SERPINA1), secretory leukocyte protease inhibitor (SLPI), and elafin (PI3), which maintain protease-antiprotease balance. Dysregulation of ELANE contributes to excessive matrix degradation and chronic inflammation, as represented by the SERPINA1/SERPINB1/ELANE/FPR1/MMP9/IL8 pathway.
Although ELANE is predominantly expressed in neutrophils, its functional analysis in the AGS gastric epithelial background provides a unique system to examine protease-driven inflammatory signaling within the gastric tumor microenvironment. This knockout model allows dissection of ELANE-mediated pathways that influence epithelial cell behavior, such as apoptosis regulation and extracellular matrix remodeling, which are relevant to cancer-associated inflammation. By comparing wild-type and ELANE-knockout AGS cells, researchers can delineate the protease??s impact on downstream effectors like MMP9 and IL8 in gastric epithelial cells.
This polyclonal knockout population is suited for applications including neutrophil biology, innate immunity, drug target screening, and genetic disease modeling of severe congenital neutropenia and cyclic neutropenia. Representative assays include Western blotting for ELANE protein levels, neutrophil elastase activity measurements, RT-qPCR for inflammatory cytokines, migration/invasion studies, co-immunoprecipitation with SERPINA1 to assess protein interactions, and flow cytometry for neutrophil-related markers. Contact Ascent Research for further information and to discuss custom experimental solutions.
