The DLG1 Knockout A-549 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of human A-549 lung carcinoma epithelial cells carrying targeted disruptions in the DLG1 gene. This polyclonal knockout pool contains a heterogeneous mixture of cells with various loss-of-function edits, providing a robust gene disruption model without clonal selection. The product serves as a versatile tool for investigating the tumor-suppressive functions of the DLG1 scaffold protein in a well-characterized pulmonary adenocarcinoma background.
The A-549 host cell line, originally isolated from a human lung adenocarcinoma, displays adherent epithelial morphology and is widely employed as a model for alveolar basal epithelial cells and type II pneumocytes. These cells retain key characteristics of lung adenocarcinoma, including epithelial markers, anchorage-dependent growth, and responsiveness to pathway modulators, making them suitable for studying cell polarity, tight junction biology, and cancer progression.
DLG1 encodes a membrane-associated guanylate kinase (MAGUK) scaffold protein that orchestrates cell junction assembly, apico-basal polarity, and signal transduction. It interacts with the cadherin-catenin complex and coordinates Hippo and Wnt pathway components. DLG1 is regulated by CDK1-mediated phosphorylation, PKC signaling, and mechanical cues via integrins; it associates with APC, PTEN, LIN7, CASK, and KIF1B. Downstream, DLG1 restrains YAP/TAZ nuclear translocation and maintains expression and localization of tight junction proteins claudin-1, occludin, and ZO-1. Loss of DLG1 disrupts junction integrity, promotes YAP/TAZ-driven transcription, and enhances RhoA GTPase activity, leading to increased proliferation and migration.
In the A-549 context, CRISPR-mediated DLG1 ablation mimics pro-invasive states observed in advanced lung adenocarcinoma. Disruption of DLG1 scaffolding function compromises epithelial barrier properties, as assessed by reduced transepithelial electrical resistance (TEER) and delocalization of ZO-1 from cell borders. Enhanced YAP/TAZ nuclear accumulation drives mesenchymal gene expression programs, linking DLG1 loss to epithelial-mesenchymal transition (EMT) and metastatic potential. This knockout model thus enables dissection of DLG1??s tumor-suppressive role in lung cancer progression, including its impact on cell cycle control and drug sensitivity.
Key research applications include elucidating DLG1??s mechanism in Hippo-mediated contact inhibition, EMT, and Wnt/??-catenin-dependent transcription; screening for modulators of junctional stability; and evaluating chemoresistance under DLG1-deficient conditions. Representative assays cover western blotting of DLG1, YAP/TAZ, and phosphorylated LATS1; immunofluorescence staining of ZO-1 and occludin; transwell migration assays; TEER measurements; MTT proliferation tests; and transcriptomic profiling via RNA-seq. For additional product details and technical support, please contact Ascent Research.
