This product is a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human HCAR2 gene in the SK-OV-3 ovarian adenocarcinoma cell line. The polyclonal format provides a genetically heterogeneous pool of cells harboring CRISPR-mediated disruptions in HCAR2, enabling loss-of-function studies without the selection bottlenecks associated with clonal isolation. This knockout model serves as a versatile tool for investigating HCAR2-mediated signaling and metabolic regulation in an epithelial cancer context.
The SK-OV-3 cell line is an established epithelial model derived from the ascitic fluid of a patient with ovarian adenocarcinoma. Widely employed in oncology research, SK-OV-3 cells recapitulate key features of high-grade serous ovarian carcinoma, including aberrant cell proliferation, migration, and resistance to apoptosis. Their utility extends to studies of tumor metabolism, drug response, and signal transduction, making them a relevant host for interrogating the role of metabolite-sensing receptors such as HCAR2.
HCAR2 encodes a Gi/o-coupled receptor that is endogenously activated by niacin (nicotinic acid), the short-chain fatty acid butyrate, and the ketone body ??-hydroxybutyrate. Upon ligand binding, HCAR2 couples to G??i/o proteins to inhibit adenylyl cyclase, thereby reducing intracellular cAMP levels and attenuating protein kinase A (PKA) activity. This cascade suppresses NF-??B signaling and, in parallel, modulates the ERK1/2 pathway. Additionally, receptor activation recruits ??-arrestin-2, which can further direct downstream signaling outputs. Through these mechanisms, HCAR2 exerts anti-inflammatory and anti-lipolytic effects, while in neoplastic settings its function may shift depending on the cellular milieu.
In the SK-OV-3 background, HCAR2 knockout offers a focused platform to dissect the receptor??s contributions to ovarian cancer biology. HCAR2 has been implicated in both tumor-suppressive and tumor-promoting processes; its anti-inflammatory signaling via NF-??B inhibition may restrain tumor progression, whereas metabolic rewiring facilitated by HCAR2 could support cancer cell proliferation and survival. Disruption of HCAR2 allows researchers to assess its impact on lipid metabolism, cytokine production, and oncogenic signaling networks relevant to dyslipidemia, atherosclerosis, and inflammatory bowel disease, as well as ovarian carcinogenesis.
This polyclonal knockout model is suited for a range of experimental workflows, including Western blotting and RT-qPCR for confirming loss of HCAR2 expression, cAMP accumulation assays to verify Gi-coupled signaling, and NF-??B luciferase reporter assays to monitor downstream transcriptional responses. Functional studies may incorporate cell viability and apoptosis assays under nutrient-modulated conditions, migration and invasion assays to evaluate metastatic potential, and Seahorse metabolic flux analysis to probe metabolic reprogramming. Additional applications encompass drug target validation for niacin/butyrate-based interventions and anti-inflammatory pathway dissection. For further information or to discuss custom applications, please contact Ascent Research.
