The Ptger3 Knockout RAW 264.7 Cell Line is a CRISPR/Cas9-edited knockout cell line that provides constitutive disruption of the Ptger3 gene in the RAW 264.7 mouse macrophage background. This product is supplied as a validated loss-of-function model, enabling researchers to study EP3 receptor signaling without the variability of transient knockdowns or the need for in-house genome editing. It is suitable for a wide range of functional assays in immunology and cancer biology.
RAW 264.7 cells are a well-established murine macrophage line derived from an Abelson murine leukemia virus-induced tumor in a male BALB/c mouse. They are widely employed as a model for monocyte/macrophage biology due to their robust phagocytic activity, cytokine secretion profiles, and responsiveness to Toll-like receptor ligands and inflammatory cytokines. This host background provides a physiologically relevant context for investigating prostaglandin signaling in innate immunity.
Ptger3 encodes the EP3 receptor, a G protein-coupled receptor that primarily activates Gi/o proteins, leading to inhibition of adenylate cyclase and decreased cAMP synthesis. This attenuates PKA/CREB signaling while engaging ??-arrestin-2-dependent pathways and cross-talk with PI3K/AKT and ERK1/2 cascades. Receptor activity is modulated by GRK2 and RGS proteins. In macrophages, EP3 expression is induced by PGE2 itself as well as by upstream pro-inflammatory mediators such as IL-1??, TNF-??, and LPS, often via NF-??B. Downstream, EP3 regulates effectors including cAMP, PKA, COX-2, RhoA, and transcriptional programs that control inflammatory cytokine production and phagocytosis.
In RAW 264.7 cells, Ptger3 knockout abrogates Gi-mediated inhibitory tone on adenylate cyclase, thereby disrupting PGE2-dependent modulation of macrophage function. This model is instrumental for deciphering EP3??s role in macrophage polarization, migration, and the balance between pro- and anti-inflammatory cytokine outputs. It provides a defined system to study the contribution of EP3 to diseases such as rheumatoid arthritis, asthma, colorectal cancer, atherosclerosis, and preterm labor.
Typical applications include measurement of cAMP levels, Western blotting for downstream effectors (e.g., phospho-ERK1/2, phospho-AKT), RT-qPCR for gene expression changes, ELISA-based cytokine profiling, phagocytosis and transwell migration assays, and flow cytometric analysis of surface markers. The line is also suited for NF-??B reporter studies and co-culture assays modeling the tumor microenvironment. For technical support and ordering details, please contact Ascent Research.
