The GPR180 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population derived from the HT29 human colorectal adenocarcinoma cell line, featuring targeted disruption of the GPR180 gene. This loss-of-function model enables systematic investigation of GPR180-dependent signaling and cellular phenotypes without clonal selection artifacts, providing a robust tool for functional genomics and cancer biology research.
HT29 is a widely used colorectal adenocarcinoma cell line originating from a female patient, characterized by wild-type TP53 status and the capacity for enterocytic differentiation under appropriate culture conditions. These epithelial cells retain features of intestinal barrier function, absorptive and secretory activities, making them a relevant model for colorectal tumor biology and differentiation studies. Their ability to form polarized monolayers and undergo differentiation further enhances their utility in studying cell?Cmatrix interactions and epithelial homeostasis.
GPR180 encodes a G-protein coupled receptor-like protein that integrates extracellular cues from EGF and integrin-mediated adhesion to regulate intracellular signaling. It acts upstream of the MAPK/ERK cascade and PI3K/AKT/mTOR pathway, with downstream effectors including ERK1/2, AKT, paxillin, and focal adhesion kinase (FAK). GPR180 functionally interacts with integrin ??1, talin, vinculin, and paxillin, linking receptor-mediated signals to cytoskeletal dynamics and focal adhesion turnover. Within the canonical EGFR pathway, GPR180 contributes to signal propagation through GRB2, SOS, RAS, RAF, MEK, and ERK, while also intersecting with PI3K, AKT, mTOR, and SRC-dependent networks, thereby coordinating adhesion, migration, and proliferation.
Disruption of GPR180 in HT29 cells attenuates activation of both MAPK/ERK and PI3K/AKT pathways, leading to impaired cell adhesion, reduced migration, and decreased proliferation. Consequently, the tumorigenic potential of these colorectal cancer cells is diminished, underscoring the role of GPR180 in maintaining the malignant phenotype. This polyclonal knockout model provides a physiologically relevant platform to dissect the molecular mechanisms by which GPR180 supports tumor progression and metastasis, offering insights into the interplay between adhesion signaling and oncogenic pathways.
The GPR180 Knockout HT29 Polyclonal Cells are ideally suited for a range of functional assays, including Western blot analysis of phospho-ERK and phospho-AKT, RT-qPCR validation of downstream targets, wound-healing and transwell invasion assays to quantify migration and invasion, cell adhesion assays, immunofluorescence localization of focal adhesion components, and flow cytometric evaluation of proliferation markers. These cells support colorectal cancer research, studies of cell migration and invasion, tumor progression modeling, drug target discovery, and functional genomics. For further information or to discuss custom applications, please contact Ascent Research.
