Dcps Knockout RAW 264.7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the RAW 264.7 mouse macrophage cell line. These cells harbor a targeted disruption of the Dcps gene, encoding the scavenger decapping enzyme essential for complete 5???3?? mRNA decay. The polyclonal format provides a heterogeneous knockout pool, suitable for pooled functional screens and bulk assays without single-cell cloning. This tool facilitates research into Dcps-dependent cap metabolism and its impact on macrophage biology.
RAW 264.7 cells, originating from BALB/c mice, serve as a classic monocyte/macrophage model characterized by robust phagocytosis, inflammatory cytokine secretion, and antigen presentation capabilities. They respond to TLR ligands, cytokines, and microbial stimuli, making them ideal for innate immunity and host-pathogen studies. Their functional versatility enables investigation of gene roles in polarization, oxidative burst, and immune signaling.
Dcps operates downstream of Dcp2-mediated decapping in the 5???3?? mRNA degradation pathway. Following deadenylation by the CCR4-NOT or PAN2-PAN3 complexes and cap removal by the Dcp1-Dcp2 decapping enzyme, Dcps hydrolyzes the m7GDP cap to m7GMP and phosphate. This reaction exposes a 5?? monophosphate that is a required substrate for processive Xrn1 exonuclease digestion. Dcps associates with P-bodies and interacts with decay factors including Dcp1a, Edc4, Lsm4, PatL1, DDX6, and EDC3, contributing to miRNA-mediated silencing and RNA quality control.
In macrophages, dynamic mRNA turnover underpins rapid responses to pathogens and inflammatory cues. Dcps deficiency in RAW 264.7 cells enables dissection of how cap scavenging activity affects cytokine output, phagocytosis, and polarization. Links to intellectual disability syndromes, viral infections, and cancer further underscore the model’s value for exploring mRNA decay in innate immunity. This knockout system offers a physiologically relevant setting to study the immunomodulatory consequences of impaired cap metabolism.
Example applications include Western blotting for Dcps and interacting proteins, RT-qPCR for mRNA half-life measurements, RNA-seq for transcriptome-wide profiling, and cap hydrolysis assays. P-body visualization by immunofluorescence, phagocytosis assays, cytokine ELISAs, and flow cytometry for macrophage markers enable functional phenotyping. For further details, contact Ascent Research.
