The GPNMB Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the GPNMB gene in the HT29 human colorectal adenocarcinoma cell line. This loss-of-function model enables systematic study of GPNMB-dependent processes without residual protein interference, with the polyclonal format offering a heterogeneous gene disruption background suitable for population-level analyses.
HT29 is an epithelial colorectal adenocarcinoma cell line derived from a primary colon adenocarcinoma of a 44-year-old Caucasian female. It retains enterocytic differentiation capacity and forms polarized monolayers, making it a well-established model for colorectal cancer research. The cell line harbors mutations in APC, p53, and other oncogenic drivers that complement GPNMB-mediated signaling studies.
GPNMB is a transmembrane glycoprotein that, upon binding integrins ??v??3 and ??5??1 or CD44, activates PI3K/AKT and ERK/MAPK signaling, leading to phosphorylation of AKT, ERK1/2, and downstream effectors such as NF-??B, GSK3??, MMP-2, and MMP-9. Shedding by ADAM10/ADAM17 generates a soluble ectodomain that modulates intercellular communication. Upstream regulators TGF-??, TNF-??, IFN-??, and MITF drive expression under hypoxic conditions. In colorectal cancer, GPNMB promotes tumor progression and immunosuppression via these pathways.
In HT29 colorectal cancer cells, GPNMB loss disrupts key oncogenic signaling cascades, allowing investigators to dissect its role in AKT- and ERK-dependent survival, migration, and immune modulation. The HT29 background, with constitutive Wnt/??-catenin and NF-??B activity, provides a physiologically relevant system for probing GPNMB?Cintegrin?CCD44 crosstalk and its impact on tumor cell plasticity and microenvironment interactions.
This knockout model is suited for western blot analysis of phospho-AKT/ERK, RT-qPCR quantification of MMP gene expression, and migration/invasion assays. Flow cytometry enables assessment of integrin ??v??3 and CD44 surface levels, and co-immunoprecipitation confirms GPNMB?CCD44 binding. Proliferation and apoptosis assays under drug treatment reveal functional consequences, while RNA-seq captures global pathway changes. Applications include colorectal cancer invasion research, tumor-immune interaction studies, and drug resistance screening. For additional information, contact Ascent Research.
