GPRIN3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. These cells carry a targeted disruption of the GPRIN3 gene, generating a loss-of-function model for investigating the roles of GPRIN3 in intestinal epithelial biology, cancer, and cytoskeletal dynamics. The polyclonal format provides a heterogeneous population of edited cells, suitable for studying gene function without clonal artifacts. This product is designed for researchers studying G protein-coupled receptor signaling, actin cytoskeleton reorganization, and cell adhesion and migration. The knockout model enables dissection of GPRIN3-dependent mechanisms in a relevant cancer cell context.
HT29 is a widely used human female colorectal adenocarcinoma cell line established in 1964 from a primary tumor. These adherent epithelial cells retain features of intestinal epithelium, including expression of characteristic markers and the ability to form polarized monolayers. HT29 cells can undergo enterocytic differentiation when treated with agents such as butyrate, making them a versatile model for colorectal cancer progression and intestinal differentiation studies. Their tumorigenic properties and well-characterized signaling pathways provide a robust platform for functional analysis of genes implicated in cancer metastasis and cytoskeletal regulation.
GPRIN3 encodes a G protein-regulated inducer of neurite outgrowth that functions as a scaffold protein linking GPCR signaling to actin dynamics. Upstream, GPRIN3 is regulated by G protein alpha subunits GNAI and GNAS, neurotrophic factors such as NGF and BDNF, cAMP, and Rho GTPases. Downstream, it activates Rac1 and RhoA, leading to modulation of focal adhesion kinase, cofilin, and the actin cytoskeleton. GPRIN3 also interacts with tubulin, actin, and integrins. The canonical pathway involves GNAI/GNAS ?? GPRIN3 ?? Rac1/Cdc42 ?? PAK ?? LIMK ?? cofilin, ultimately driving actin filament reorganization. This network positions GPRIN3 as a critical node in cell adhesion, migration, and morphological changes.
In HT29 cells, GPRIN3 likely regulates adhesion and migration through cytoskeletal mechanisms analogous to its role in neurons. Disruption of GPRIN3 in this colorectal cancer model permits detailed study of GPCR-mediated actin remodeling, integrin signaling, and cell motility??processes central to tumor invasion and metastasis. The polyclonal knockout population enables assessment of GPRIN3??s contribution to the morphological plasticity of HT29 cells, including their capacity to form polarized monolayers and to undergo epithelial-to-mesenchymal transition. This model is particularly relevant for exploring links between neuronal differentiation pathways and neuroendocrine features in colon cancer.
Researchers can use these GPRIN3 Knockout HT29 Polyclonal Cells in a range of functional assays to dissect signaling mechanisms and evaluate therapeutic interventions. Typical applications include wound healing and transwell migration/invasion assays, immunofluorescence for actin filaments, co-immunoprecipitation with G protein subunits, western blotting for Rho GTPase activation, RT-qPCR for GPRIN3 expression, and cell adhesion assays. These knockout cells facilitate colon cancer metastasis modeling, GPCR signaling analysis, and screening of GPRIN3 modulators. They also support studies of neuroendocrine differentiation in colon cancer. For further information, please contact Ascent Research.
