The DLG5 Knockout NCI-H1299 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated through CRISPR/Cas9-mediated disruption of the DLG5 gene in the NCI-H1299 human lung adenocarcinoma cell line. This heterogeneous pool provides a versatile loss-of-function model for investigating DLG5-dependent cellular processes without the selection of a single clonal isolate.
NCI-H1299 is a widely used non-small cell lung cancer (NSCLC) cell line derived from the metastatic lymph node of a 43-year-old male with lung adenocarcinoma. These adherent epithelial cells are p53-deficient, which abrogates canonical p53-mediated responses and makes the line particularly suitable for studying apoptosis, proliferation, and metastasis independent of p53 status.
DLG5 encodes a membrane-associated guanylate kinase scaffold protein that localizes to tight junctions and the apical cortex, where it integrates epithelial polarity with Hippo pathway regulation. It interacts with polarity complex components PALS1, PATJ, and CRB3, and directly associates with Hippo kinases MST1/2 and LATS1/2. Through these interactions, DLG5 modulates YAP/TAZ phosphorylation and subcellular localization, thereby influencing TEAD-mediated transcription. DLG5 also participates in Wnt signaling by binding to CTNNB1 (??-catenin), linking cell-cell adhesion to transcriptional control.
In NCI-H1299 cells, DLG5 knockout disrupts junctional integrity and apical-basal polarity, leading to altered YAP/TAZ activity and a more migratory, invasive phenotype. This perturbation is particularly informative in a p53-null background, enabling dissection of Hippo and polarity pathways without confounding p53-dependent effects. The model thus offers a clean system for studying mechanisms of epithelial-mesenchymal transition (EMT) and metastatic progression in lung adenocarcinoma.
These polyclonal knockout cells are suitable for a broad range of assays. Western blotting can assess YAP/TAZ phosphorylation status and expression of tight junction proteins such as CLDN1 and OCLN, while immunofluorescence enables visualization of junctional markers like ZO-1 and occludin. Transwell migration and invasion assays, RhoA activity measurements, and RNA sequencing of Hippo target genes can characterize functional and transcriptional changes. Co-immunoprecipitation can probe altered protein interactions within the DLG5 network. The model supports detailed investigation of EMT, cancer cell motility, and Hippo/Wnt crosstalk in lung adenocarcinoma. For further information, please contact Ascent Research.
