The ADGRG1 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human A-549 lung adenocarcinoma line. The ADGRG1 gene has been disrupted to create a loss-of-function model, resulting in a heterogeneous pool of edited cells lacking functional GPR56 protein. This polyclonal knockout stock is supplied as a convenient and robust tool for studying adhesion GPCR biology in cancer, avoiding the biases of clonal selection.
The parental A-549 cell line is a well-established model of human lung adenocarcinoma, carrying a KRAS G12S mutation and wild-type p53. These epithelial cells are widely employed in non-small cell lung cancer research, displaying characteristic tumorigenic features such as xenograft tumor formation and in vitro migratory capacity. The stable genetic background and extensive characterization of A-549 cells make them an ideal host for gene-editing studies focused on adhesion and metastasis.
ADGRG1 encodes the adhesion GPCR GPR56, which binds collagen III and transglutaminase 2 to activate G??12/13-mediated RhoA signaling. This pathway stimulates ROCK, NF-??B, and MMP2, while also promoting VEGF expression, collectively regulating cell adhesion, migration, and invasion. GPR56 signaling exhibits crosstalk with integrins and the Wnt/??-catenin pathway, and transglutaminase 2 functions as both a ligand and scaffold. Disruption of ADGRG1 therefore abolishes these receptor-mediated signals, offering a clean background to dissect GPR56-dependent molecular mechanisms.
In A-549 lung adenocarcinoma cells with oncogenic KRAS, ADGRG1 expression likely contributes to enhanced matrix adhesion and metastatic behavior. Knockout of this receptor disrupts GPR56-collagen III interactions and downstream RhoA/ROCK signaling, potentially attenuating pathways that drive tumor progression. Given its relevance in non-small cell lung cancer and other malignancies, this model enables precise investigation of GPR56’s role in adhesion dynamics and signaling within a clinically relevant genetic background.
The polyclonal knockout population is ideally suited for transwell migration and adhesion assays to quantify metastatic potential, as well as for western blotting, phospho-signaling analysis, and RNA-seq to map RhoA/ROCK pathway alterations. Drug screening and target validation studies benefit from the pooled format’s assessment of GPR56 dependence. Apoptosis and proliferation assays further characterize phenotypic consequences, while flow cytometry and RT-qPCR confirm gene disruption. For product inquiries, contact Ascent Research.
