The HCAR2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human cell line, targeting the HCAR2 gene. This product provides a heterogeneous pool of cells carrying diverse loss-of-function mutations in HCAR2, enabling robust functional genomics studies of this G protein-coupled receptor. The polyclonal format preserves population-level representation and is ideal for pooled screening and pathway dissection without clonal bias. By disrupting HCAR2, the cells serve as a versatile loss-of-function model for investigating receptor-mediated signaling and its physiological consequences.
The HAP1 cell line is a near-haploid human myeloid cell line originally derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype simplifies gene editing and knockout generation, making it a widely adopted model for CRISPR-based functional genomics, drug target validation, and signaling pathway analysis. HAP1 cells retain key signaling networks relevant to hematological and inflammatory research, providing a genetically stable and accessible platform for mechanistic studies. Their near-haploid state also facilitates high-efficiency genome engineering and subsequent phenotype screening.
HCAR2 (also known as GPR109A) encodes a Gi/o-coupled receptor that responds to the endogenous ligands niacin, butyrate, and beta-hydroxybutyrate. Upon activation, HCAR2 couples to heterotrimeric Gi/o proteins, inhibiting adenylate cyclase and thereby reducing intracellular cAMP levels. The consequent decrease in PKA activity attenuates hormone-sensitive lipase-mediated lipolysis in adipocytes and dampens NF-??B-driven pro-inflammatory transcriptional programs in immune cells. Additionally, HCAR2 engagement modulates the MAPK/ERK and Akt signaling pathways, influencing cell proliferation and survival. The receptor exhibits ??-arrestin recruitment, which may contribute to desensitization and downstream signaling events.
In the HAP1 background, HCAR2 knockout provides a genetically tractable model to dissect the receptor??s signaling functions in a haematopoietic context. Although HCAR2 is well characterized in adipocytes, its expression in myeloid and other immune cells underscores its role in modulating inflammatory responses, as seen in atherosclerosis, colitis, and metabolic syndrome. The knockout cells enable systematic investigation of HCAR2-dependent signaling networks, including Gi/o-mediated cAMP suppression, ERK phosphorylation, and NF-??B activity, in a homogenous cell system amenable to high-throughput genetic and pharmacological manipulation.
These polyclonal knockout cells are well-suited for functional genomics screens, GPCR signal transduction studies, and anti-inflammatory compound profiling. Researchers can assess HCAR2-dependent changes in cAMP levels, MAPK/ERK and Akt phosphorylation, NF-??B reporter activity, and downstream gene expression by RT-qPCR. The cells also support immunofluorescence and flow cytometry-based experiments to monitor receptor surface expression or signal pathway activation. The HCAR2 Knockout HAP1 Polyclonal Cells thus offer a flexible platform for mechanistic dissection and drug discovery across inflammation, metabolism, and cardiovascular research areas. For further details or technical consultation, please reach out to Ascent Research.
