The GNG12 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the GNG12 gene in the human colorectal adenocarcinoma cell line HT29. This heterogeneous cell pool contains a variety of CRISPR-mediated modifications at the GNG12 locus, yielding a loss-of-function model for studying heterotrimeric G protein signaling. The polyclonal format provides a more representative biological replicate than monoclonal lines, reducing clonal selection artifacts and enabling robust functional genomics studies in an epithelial background.
HT29 cells originate from a human colorectal adenocarcinoma and serve as a well-characterized intestinal epithelial model. They are extensively used in cancer research for their ability to recapitulate key aspects of colorectal tumorigenesis, including ligand-dependent activation of the MAPK/ERK and PI3K-AKT pathways. This adherent cell line retains functional GPCR expression and downstream kinases, making it an ideal system for investigating G?¦?-mediated signal transduction in the context of colorectal cancer.
GNG12 encodes the gamma 12 subunit of heterotrimeric G proteins, which pairs with G?? subunits (GNB1?C5) to form the G?¦? dimer. Following GPCR activation??by chemokine receptors, lysophosphatidic acid receptors, or indirect cross-talk with receptor tyrosine kinases??the G?¦? dissociates from G?? and directly stimulates effectors including phosphoinositide 3-kinase (PI3K), phospholipase C ?? (PLC??), adenylyl cyclase isoforms, and G-protein-gated inwardly rectifying potassium (GIRK) channels. In turn, this triggers AKT phosphorylation, calcium flux, and activation of the Raf/MEK/ERK1/2 signaling cascade. The dimer is also regulated by RGS proteins and scaffold factors such as AGS3.
In HT29 colorectal adenocarcinoma cells, G?¦? signals are critical for promoting proliferation, migration, and invasion. Disruption of GNG12 abolishes functional G?¦? dimers, uncoupling GPCRs from key oncogenic pathways. This knockout model allows researchers to dissect the G?¦?-dependent components of colorectal cancer progression, evaluate compensatory signaling rewiring, and assess the therapeutic potential of G?¦?-targeted inhibitors in a disease-relevant system. It is particularly useful for studying chemokine-driven metastatic mechanisms.
The GNG12 Knockout HT29 Polyclonal Cells support a variety of assay formats, including MTT or BrdU proliferation studies, transwell migration and invasion assays, calcium flux analysis, and cAMP accumulation measurements. Pathway activation can be quantified by western blotting for phospho-ERK and phospho-AKT. The cells are amenable to high-throughput screening for G?¦? inhibitors and to RNA-seq transcriptional profiling. For further information, please contact Ascent Research.
