The HCAR2 Knockout A-549 Polyclonal Cells product offers a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the human HCAR2 gene in the A-549 lung adenocarcinoma cell line. This heterogeneous polyclonal pool provides a robust loss-of-function model, avoiding clonal selection biases while maintaining efficient gene knockout across the population for reliable functional studies.
The A-549 host cell line is a well-established human alveolar basal epithelial adenocarcinoma model, originally isolated from a 58-year-old male patient. Widely used in cancer biology, these cells exhibit key epithelial characteristics and are amenable to genetic manipulation, making them an ideal platform for generating knockout derivatives and performing downstream phenotypic assays.
HCAR2 encodes the hydroxycarboxylic acid receptor 2, a G protein-coupled receptor activated by niacin, butyrate, beta-hydroxybutyrate, and free fatty acids. Upon ligand binding, HCAR2 couples to Gi/o alpha subunits, inhibiting adenylyl cyclase and reducing intracellular cAMP levels. This signaling cascade involves downstream factors including PKA and AMPK, and converges on pathways such as MAPK/ERK and NF-kB, with HCAR2 activation leading to NF-kB inhibition. The receptor also interacts with beta-arrestin and G protein-coupled receptor kinases, and regulates the ABCA1 transporter, collectively mediating anti-lipolytic and anti-inflammatory effects.
In the A-549 lung adenocarcinoma context, HCAR2 knockout cells enable dissection of receptor functions in cancer-related processes such as lipid metabolism, inflammatory signaling, and cell migration. Since HCAR2 ligands like butyrate are gut microbiome-derived and can reach systemic circulation, these cells provide a relevant model for studying how dietary metabolites influence lung tumor biology. Moreover, the A-549 background expresses requisite GPCR signaling components, facilitating investigation of HCAR2-mediated anti-inflammatory pathways and their interplay with oncogenic networks.
These polyclonal knockout cells are suitable for a range of assays, including cAMP accumulation measurements to assess Gi-mediated signaling, western blot detection of phospho-AMPK to monitor metabolic pathway engagement, and RT-qPCR analysis of inflammatory cytokines such as IL-6 and TNF-alpha. Additional applications include lipid uptake assays and migration studies, supporting research in lipid metabolism, anti-inflammatory drug development, and cancer progression. For further information or to explore potential collaborations, please contact Ascent Research.
