HCAR2 Knockout MCF-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the MCF-7 human breast adenocarcinoma cell line. The product provides a heterogeneous pool of cells harboring targeted disruptions of the HCAR2 gene, enabling loss-of-function studies without clonal isolation. This polyclonal format is well-suited for experiments requiring bulk population-level analysis of HCAR2-dependent phenotypes, while minimizing the confounding effects of single-cell clonal variation. The knockout model supports robust interrogation of HCAR2-mediated signaling in a well-characterized estrogen-responsive breast cancer background.
The parental MCF-7 cell line was originally established from a pleural effusion of a 69-year-old patient with invasive breast ductal carcinoma. MCF-7 cells maintain an epithelial morphology and are positive for both estrogen receptor (ER) and progesterone receptor (PR), with wild-type p53 status. These characteristics render MCF-7 a widely used model for hormone-responsive breast cancer, particularly in studies of ER-dependent proliferation, transcriptional regulation, and endocrine therapy resistance. The line??s genetic stability and extensive molecular characterization provide a reliable context for gene-editing experiments.
HCAR2 is a Gi/o-coupled receptor activated by the endogenous metabolite butyrate and the pharmacologic agent nicotinic acid (niacin). Ligand binding leads to G??i/o-mediated inhibition of adenylate cyclase, reducing intracellular cAMP levels and protein kinase A (PKA) activity. Downstream, this attenuates hormone-sensitive lipase activation and perilipin phosphorylation, suppressing lipolysis. Additionally, HCAR2 signaling engages ??-arrestin and G?¦? subunits, modulating ERK1/2 and NF-??B to promote anti-inflammatory responses such as IL-10 expression. Receptor expression is induced by PPAR??, and activation by butyrate or ??-hydroxybutyrate links metabolic states to immune and proliferative control.
In the context of MCF-7 breast cancer cells, HCAR2 knockout disrupts the normal inhibitory tone on adenylate cyclase, leading to derepressed cAMP/PKA signaling upon ligand challenge. This alteration can impact cell cycle progression, apoptosis, and inflammatory cytokine secretion, as HCAR2 has been implicated in modulating the balance between survival and anti-inflammatory gene programs. Because MCF-7 cells are responsive to estrogen and butyrate/nicotinic acid, the polyclonal knockout population allows researchers to dissect how HCAR2 loss influences ER-driven oncogenic pathways and metabolic signaling crosstalk. Potential phenotypic consequences include altered proliferation rates, migratory capacity, and sensitivity to anti-cancer agents, making this model valuable for investigating the role of metabolite-sensing GPCRs in breast tumor biology.
This HCAR2 knockout product is intended for a broad spectrum of research applications. Typical uses include validation of small-molecule agonists or antagonists targeting HCAR2, mechanistic studies of GPCR signal transduction, and functional assays measuring cAMP accumulation, phospho-ERK status, or IL-10 production following butyrate or niacin treatment. Standard readouts may employ Western blotting for downstream targets such as PKA substrates or phospho-ERK1/2, cAMP ELISAs, RT-qPCR for gene expression changes, MTT cell viability assays, and NF-??B luciferase reporter systems. The polyclonal nature makes it suitable for pooled functional screens and bulk pharmacodynamic assessments. For further technical information or to discuss custom cell engineering needs, please contact Ascent Research.
