This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line, engineered to harbor a targeted disruption of the DLG5 gene. This heterogeneous pool of edited cells enables loss-of-function studies without the limitations of single-cell-derived clones, providing a physiologically relevant model system.
The AGS parental line, isolated from a gastric adenocarcinoma, is a widely used epithelial model that retains key characteristics of gastric cancer cells, including anchorage-independent growth and aberrations in cell adhesion and signaling pathways. Its epithelial origin and tumorigenic background make it particularly suited for investigating genes involved in cell polarity, junction dynamics, and carcinogenesis.
DLG5 encodes a membrane-associated guanylate kinase scaffolding protein integral to the formation and maintenance of apical junction complexes. It physically interacts with PALS1, PATJ, and CRB3 to establish apicobasal polarity, while also binding ??-catenin and cadherins at adherens junctions. Functionally, DLG5 promotes Hippo pathway activation by facilitating MST1/2 and LATS1/2 kinase activity, resulting in phosphorylation and cytoplasmic sequestration of the transcriptional co-activators YAP and TAZ. This restricts their nuclear entry and the expression of pro-proliferative and pro-migratory target genes. DLG5 activity is regulated by upstream cues including E-cadherin engagement, Wnt ligands, cell density, and AMPK, positioning it as a critical integrator of mechanical and biochemical signals at cell?Ccell contacts.
In the AGS background, disruption of DLG5 is predicted to dismantle tight junction integrity, impair YAP/TAZ restriction, and dysregulate Wnt/??-catenin signaling, potentially driving enhanced proliferation, migration, and loss of contact inhibition??hallmarks of gastric cancer progression. The polyclonal knockout population thus recapitulates key aspects of tumor suppressor loss and junctional disassembly seen in gastric and colorectal cancers, as well as in inflammatory bowel diseases such as Crohn??s disease, where DLG5 polymorphisms have been implicated.
Researchers can utilize this polyclonal knockout model for multifaceted investigations: western blotting and immunofluorescence to assess changes in junctional proteins (claudins, occludin, ZO-1) and polarity markers; phospho-YAP analysis and subcellular fractionation to quantitate Hippo pathway output; co-immunoprecipitation to interrogate DLG5 interactions with PALS1, PATJ, or ??-catenin; transepithelial electrical resistance (TEER) assays to measure barrier function; and migration/invasion assays to evaluate metastatic potential. Transcriptomic profiling via RNA-seq or targeted RT-qPCR can identify downstream transcriptional consequences, while apoptosis assays reveal effects on cell survival. These applications make the cell population a versatile tool for dissecting DLG5-dependent mechanisms in epithelial homeostasis and disease. For additional information or technical assistance, please contact Ascent Research.
