The GPR108 Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population derived from the human colorectal adenocarcinoma cell line HT29, engineered for loss-of-function studies of the G protein-coupled receptor 108 (GPR108). This polyclonal knockout model provides a mixture of cells harboring diverse editing events at the GPR108 locus, enabling robust analysis of gene function without clonal selection bias. The product is designed for researchers investigating innate immune signaling, NF-??B pathway regulation, and type I interferon responses in the context of colorectal cancer and inflammatory diseases.
The parental HT29 cell line originated from a primary colorectal adenocarcinoma and is extensively employed as an intestinal epithelial model. These cells exhibit characteristics of absorptive enterocytes and retain the ability to form polarized monolayers, making them valuable for studying drug transport, barrier function, and epithelial-to-mesenchymal transition (EMT). HT29 cells also express key innate immune signaling components, allowing dissection of host defense mechanisms in a colorectal cancer background.
GPR108 functions as a negative regulator of the type I interferon response by directly interacting with TANK-binding kinase 1 (TBK1) and inhibiting its phosphorylation, thereby suppressing downstream activation of interferon regulatory factor 3 (IRF3) and expression of IFN-?? and interferon-stimulated genes (ISGs). Additionally, GPR108 may modulate NF-??B signaling through interactions with I??B kinase epsilon (IKK??) and the mitochondrial antiviral-signaling protein (MAVS). Upstream activation by inflammatory cytokines such as TNF-?? and IL-1??, Toll-like receptor ligands like lipopolysaccharide, or viral stimuli triggers canonical signaling cascades involving TBK1, IRF3, p65, and STAT1. The knockout of GPR108 is anticipated to relieve this inhibitory constraint, potentially enhancing innate immune pathway activation in HT29 cells.
In the context of HT29 colorectal cancer cells, disruption of GPR108 offers a unique platform to explore the interplay between oncogenic signaling and innate immunity. Colorectal tumors often evade immune surveillance; thus, by eliminating a negative regulator of interferon and NF-??B responses, the GPR108 knockout model can help elucidate mechanisms governing tumor-cell-intrinsic immune signaling. This is particularly relevant for investigating how colorectal cancer cells respond to microbial stimuli and inflammatory mediators, potentially revealing therapeutic vulnerabilities or immunomodulatory strategies.
The GPR108 Knockout HT29 Polyclonal Cells are suited for diverse experimental approaches including Western blotting for phospho-TBK1 and IRF3, RT-qPCR for IFN-?? and ISGs, dual luciferase reporter assays for NF-??B and ISRE activity, co-immunoprecipitation of GPR108 interacting partners, and cytokine ELISA profiling. These cells enable host-pathogen interaction studies, GPCR functional characterization, and drug target validation for inflammatory diseases and colorectal cancer. They provide a powerful tool for interferon pathway screening and innate immune signal transduction research. For further information, please contact Ascent Research.
