The HCAR2 Knockout HeLa Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population based on the HeLa cell line, designed to disrupt the HCAR2 gene encoding the hydroxycarboxylic acid receptor 2 (HCAR2, also known as GPR109A). This heterogeneous loss-of-function model, generated through CRISPR/Cas9-mediated gene disruption, provides a pooled population of cells carrying various mutations in the target locus, enabling rigorous assessment of HCAR2-dependent signaling pathways without the biases associated with single clones.
The parental HeLa line is an immortalized human cervical epithelial adenocarcinoma cell line originally derived from a 31-year-old patient and is positive for human papillomavirus type 18 (HPV18). HeLa cells are a foundational model in cancer biology, cell cycle research, and signal transduction studies, offering robust growth characteristics and well-characterized molecular pathways. Their sustained use in both basic and translational research makes them an ideal host for gene-editing approaches, particularly for exploring receptor function in a cancer context.
HCAR2 functions as a Gi/o protein-coupled receptor activated by ligands including nicotinic acid (niacin), butyrate, and beta-hydroxybutyrate, as well as PPARG agonists. Upon activation, HCAR2 couples to G-alpha-i subunits to inhibit adenylyl cyclase, reducing intracellular cAMP. This attenuates cAMP-dependent pathways, modulating ERK1/2 phosphorylation and suppressing transcription factors NF-??B and CREB. The receptor interacts with beta-arrestins and GRK2 for desensitization and internalization. Consequently, HCAR2 activation downregulates pro-inflammatory cytokines IL-6 and TNF-alpha and promotes adiponectin secretion, underscoring its role in anti-inflammatory and metabolic regulation.
In the HeLa cervical cancer background, loss of HCAR2 eliminates the cellular responses to niacin and butyrate, allowing researchers to dissect receptor-specific contributions to processes often dysregulated in cancer, such as proliferation, migration, and inflammatory signaling. This knockout model is particularly valuable for distinguishing HCAR2-dependent effects from other GPCR-mediated pathways that converge on cAMP and NF-??B. Because HeLa cells inherently exhibit altered cell cycle control and viral oncogene expression, the removal of HCAR2 provides a clean system to investigate how metabolic and anti-inflammatory signals intersect with carcinogenic drivers, potentially clarifying the receptor??s role in tumor microenvironment modulation.
Researchers can employ these knockout cells in diverse functional assays to study HCAR2 pharmacology and signaling. Typical applications include cAMP ELISA, phospho-ERK1/2 Western blotting, NF-??B luciferase reporter assays, and cytokine profiling for IL-6 and TNF-alpha secretion. Flow cytometric analysis of receptor surface expression and Transwell migration assays further enable investigation of anti-inflammatory and metabolic functions. These cells are suited for drug target validation in dyslipidemia, atherosclerosis, and inflammatory disorders, and for screening novel HCAR2 agonists or antagonists. For additional details, please contact Ascent Research.
