GPNMB Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line. This product provides a genetically disrupted GPNMB locus, enabling loss-of-function studies of the glycoprotein NMB (GPNMB) transmembrane protein. The polyclonal format ensures representation of diverse editing events across the population, offering a robust model for investigating GPNMB-dependent phenotypes without the constraints of single-cell clonal variation.
HCT 116 cells are an extensively characterized epithelial cell line originating from a human colorectal carcinoma. They harbor notable oncogenic mutations, including KRAS G13D and PIK3CA H1047R, and are deficient in mismatch repair due to MLH1 loss, conferring a microsatellite instability-high (MSI-H) phenotype. These cells are widely employed as a model for colorectal cancer studies, particularly in the context of proliferation, apoptosis, metastasis, and therapeutic response. Their adherent growth and well-defined signaling landscape make them highly amenable to gene perturbation experiments.
GPNMB encodes a type I transmembrane glycoprotein that undergoes proteolytic cleavage by ADAM10, releasing an extracellular domain (ECD). The shed ECD functions as a ligand for integrins such as ??v??3 and ??5??1 and for syndecan-4, initiating intracellular signaling cascades. Downstream, GPNMB-mediated adhesion triggers phosphorylation of focal adhesion kinase (FAK) and Src, leading to activation of AKT/mTOR and MAPK/ERK pathways. Additionally, GPNMB expression is regulated by transcription factors MITF, TGF-??1-induced SMADs, STAT3, and HIF-1??, integrating cues from the tumor microenvironment. Its signaling promotes transcription of genes including matrix metalloproteinase-9 (MMP-9), cyclin D1, and survivin, thereby enhancing cell migration, proliferation, and survival.
In HCT 116 cells, disruption of GPNMB allows dissection of its roles in colorectal carcinoma biology. Given the line??s activated KRAS and PI3K pathways, GPNMB knockout can reveal contributions to ERK and AKT signal modulation, particularly in the context of integrin-mediated adhesion and migration. Moreover, because GPNMB is implicated in epithelial-mesenchymal transition (EMT) and immunosuppression through mechanisms such as CD44 interaction and syndecan-mediated signaling, this model is valuable for probing crosstalk between oncogenic signaling and immune evasion in an MSI-high setting. The polyclonal knockout population thus provides a versatile platform for elucidating GPNMB-dependent phenotypes without clonal bias.
Typical applications include transwell migration and invasion assays to assess metastatic potential, phospho-specific immunoblotting for FAK, AKT, and ERK1/2 activation, and co-culture experiments to evaluate effects on immune cell suppression. This model also supports screening of GPNMB-targeted therapeutics, such as antibody-drug conjugates targeting the extracellular domain. Additionally, measurement of ADAM10-mediated shedding and downstream target expression by RT-qPCR or Western blot can be performed. Researchers may employ this knockout for comparative oncogene studies or osteoblast differentiation assays, given GPNMB??s dual roles. For further information, please contact Ascent Research.
