The GPR75 Knockout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, featuring targeted disruption of the GPR75 gene. This loss-of-function model provides a genetically heterogeneous pool of edited cells, enabling robust functional studies without selecting a single clonal isolate. The polyclonal format preserves biological variability while allowing researchers to assess the collective impact of GPR75 ablation on cellular phenotypes and signaling networks relevant to metabolic and inflammatory disease.
HT29 cells, isolated from a primary colon adenocarcinoma of a 44-year-old female, represent a well-characterized model of intestinal epithelial biology. These cells form polarized monolayers and can undergo differentiation under appropriate culture conditions, mimicking key aspects of the colonic epithelium. Their tumor-derived origin and capacity for polarization make them suitable for investigating oncogenic and epithelial transport processes. The combination of GPR75 knockout with HT29’s intrinsic properties creates a versatile platform for studying receptor-mediated signaling in a physiologically relevant context.
GPR75 encodes an orphan G protein-coupled receptor that functions as a receptor for the chemokine CCL5 (RANTES). Ligand engagement leads to coupling with G??i/o and G??q/11 proteins, acutely inhibiting adenylyl cyclase and reducing intracellular cAMP, while simultaneously triggering the MAPK/ERK cascade through RAF/MEK/ERK phosphorylation. Downstream effector molecules include PKA, calcium mobilization, and transcription factors such as CREB and AP-1. GPR75 also recruits ??-arrestin and is regulated by GPCR kinases (GRKs). This signaling architecture integrates energy homeostasis, inflammatory responses, and cell migration pathways.
In the HT29 intestinal epithelial context, GPR75 knockout disrupts chemokine-regulated signaling networks that control metabolism and inflammation. Since HT29 cells express cytokine and chemokine receptors and are responsive to inflammatory stimuli, ablation of GPR75 allows dissection of its specific contribution to CCL5-dependent effects on cAMP levels, MAPK activation, and gene expression. This model is particularly valuable for exploring how intestinal epithelial cells may modulate systemic energy balance and local inflammatory processes, bridging observations in metabolic tissues with gut-derived signals.
Typical research applications include obesity and type 2 diabetes research, drug target validation for anti-obesity therapeutics, chemokine signaling pathway analysis, and metabolic syndrome studies. Compatible assays encompass Western blotting for downstream targets (e.g., phospho-ERK, CREB), RT-qPCR for transcriptionally regulated genes, cAMP accumulation measurements, calcium flux assays, MAPK phospho-signaling analysis, and cell migration/invasion assays. The pooled polyclonal population facilitates population-level functional assessments. For additional information or custom knockout services, please contact Ascent Research.
