Genome-edited Cells
Ascites
The Pcyt2 Knockout RAW 264.7 Cell Line is a CRISPR/Cas9-edited macrophage model with targeted disruption of Pcyt2, the rate-limiting enzyme for phosphatidylethanolamine biosynthesis. Pcyt2 activity is regulated by SREBP1c and PPAR?? and is central to the CDP-ethanolamine pathway, affecting membrane phospholipid composition and autophagosome formation via LC3 lipidation. This knockout cell line enables investigation of phospholipid metabolism in innate immunity, autophagy, and inflammatory signaling. Key applications include phagocytosis assays, cytokine profiling, lipidomics, and LC3-based autophagy analysis, providing insights into macrophage function in metabolic disorders and lipodystrophy.
ASAP2 Knockout K562 Polyclonal Cells
Cat. No. ARG20224
PEG10 Knockout NCI-H1975 Polyclonal Cells
Cat. No. ARG16332
MGAT5 Knockout SK-HEP-1 Polyclonal Cells
Cat. No. ARG15535
NTRK3 Knockout HT29 Polyclonal Cells
Cat. No. ARG14414
ARFIP1 Knockout HT29 Polyclonal Cells
Cat. No. ARG33009
MJ
Cat. No. ARC0522
The Pcyt2 Knockout RAW 264.7 Cell Line is a CRISPR/Cas9-edited macrophage model featuring targeted disruption of the Pcyt2 gene in the Raw 264.7 murine macrophage background. This knockout cell line serves as a powerful tool for dissecting the role of the CDP-ethanolamine pathway in phospholipid metabolism and innate immune cell function. By eliminating Pcyt2 expression, researchers can investigate de novo phosphatidylethanolamine (PE) synthesis and its downstream effects on membrane biology, autophagy, and macrophage-driven processes.
Raw 264.7 cells are a widely used BALB/c-derived macrophage line that recapitulates key features of primary macrophages, including robust phagocytic activity, cytokine secretion, and responsiveness to inflammatory stimuli. They provide a genetically tractable platform for studying macrophage biology in health and disease. This immortalized line maintains the ability to polarize toward different activation states and is commonly employed in studies of infection, inflammation, and metabolic signaling.
Pcyt2 encodes CTP:phosphoethanolamine cytidylyltransferase, the rate-limiting enzyme in the CDP-ethanolamine branch of PE biosynthesis. Pcyt2 activity is transcriptionally regulated by SREBP1c and PPAR?? in response to insulin and nutritional cues. The enzyme catalyzes the formation of CDP-ethanolamine from phosphoethanolamine and CTP, which subsequently contributes to the production of PE, a major membrane phospholipid. Downstream, PE abundance directly influences autophagosome formation through LC3 lipidation, mitochondrial function, and membrane lipid remodeling. Pcyt2 also intersects with the phosphatidylethanolamine methyltransferase (PEMT) pathway and interacts with factors such as creatine kinase.
In macrophages, PE is essential for membrane curvature during phagocytosis and for the biogenesis of autophagosomes that clear intracellular pathogens and damaged organelles. The absence of Pcyt2-mediated PE synthesis can therefore compromise phagocytic capacity, autophagy flux, and inflammatory cytokine output, linking phospholipid metabolism to innate immune effector functions. This knockout model enables dissection of how lipid composition governs macrophage activation and the interplay between metabolic and immune signaling pathways.
Researchers can employ this cell line to explore PE-dependent processes in innate immunity, including phagocytosis assays, LC3 lipidation western blotting, cytokine ELISA profiling, and lipidomics analysis. It is also suitable for investigating mitochondrial membrane potential and autophagy-related signaling. Additionally, the line supports disease modeling efforts in lipodystrophy and metabolic disorders where Pcyt2 deficiency is implicated. For further information, please contact Ascent Research.
