The HCAR2 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the HCAR2 gene in the HT29 human colorectal adenocarcinoma cell line. This product provides a biologically diverse pool of edited cells, enabling robust loss-of-function analysis without the constraints of clonal selection. The polyclonal format ensures heterogeneous genetic modifications across the population, offering a representative model for studying HCAR2-dependent phenotypes in a cancer-relevant epithelial background.
The HT29 cell line is an adherent epithelial model isolated from a primary colorectal adenocarcinoma of a 44-year-old female. Widely utilized in gastrointestinal research, HT29 cells are instrumental for investigating intestinal absorption, barrier function, and colorectal cancer biology. Their ability to differentiate and form polarized monolayers under specific culture conditions makes them particularly suitable for studying epithelial transport and host?Cmicrobe interactions, thereby providing a physiologically relevant context for functional genomics studies.
HCAR2 (also designated GPR109A) encodes a Gi/o-coupled receptor stimulated by endogenous ligands such as niacin (nicotinic acid), beta-hydroxybutyrate, and the short-chain fatty acid butyrate, as well as synthetic agonists like MK-0354. Upon activation, HCAR2 triggers G??i-mediated inhibition of adenylate cyclase, leading to decreased intracellular cAMP levels and attenuated protein kinase A activity. This cascade suppresses hormone-sensitive lipase activity in adipocytes and reduces NF-kB signaling, resulting in diminished transcription of pro-inflammatory cytokines including TNF-?? and IL-6. The receptor further engages beta-arrestins and GPCR kinases to modulate signal termination and downstream effector coupling.
In the HT29 colonic epithelial context, HCAR2 knockout provides a valuable tool to dissect the receptor??s contributions to metabolic regulation and inflammation. HT29 cells endogenously express components of the NF-kB pathway and secrete cytokines, making them responsive to butyrate-mediated anti-inflammatory signals that may involve HCAR2. Disruption of HCAR2 in these cells allows researchers to investigate how the receptor modulates lipolysis-related signaling in non-adipocyte lineages, influences epithelial barrier integrity, and intersects with oncogenic pathways in colorectal cancer progression.
The HCAR2 Knockout HT29 Polyclonal Cells are suited for a broad range of applications, including colorectal cancer metabolism studies, anti-inflammatory drug screening, and GPCR functional assays. Representative techniques include cAMP accumulation assays to quantify Gi signaling, NF-kB reporter assays to monitor transcriptional responses, cytokine ELISA for TNF-?? and IL-6, as well as cell viability and migration/invasion assays to assess cancer cell behavior. For additional technical information, please contact Ascent Research.
