The Dusp1 Knockout RAW 264.7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population featuring targeted disruption of the Dusp1 gene in the RAW 264.7 murine macrophage cell line. This loss-of-function model enables investigation of DUSP1 (MKP-1) function in inflammatory signaling, providing a robust system that avoids the selective biases of single-cell cloning.
RAW 264.7 cells, derived from an Abelson virus-transformed BALB/c mouse macrophage, are a widely used model for innate immunity. They exhibit robust phagocytic activity and produce numerous cytokines in response to TLR stimulation, particularly via TLR4 ligands such as lipopolysaccharide (LPS), and are a standard host for macrophage biology studies.
DUSP1 is a nuclear dual-specificity phosphatase that dephosphorylates and inactivates ERK1/2, JNK, and p38 MAP kinases, acting as a critical negative regulator of MAPK cascades. Its expression is induced by TLR4 ligands, TNF-??, IL-1??, and reactive oxygen species through AP-1 and NF-??B transcription factors. DUSP1 functions downstream of MAP3Ks (Raf, ASK1, TAK1) and MAP2Ks (MEK1/2, MKK4/7, MKK3/6) and directly terminates signaling by removing phosphates from the activation loop of terminal MAPKs.
In RAW 264.7 macrophages, Dusp1 knockout results in sustained MAPK activation and exaggerated production of pro-inflammatory cytokines like TNF-?? and IL-6 upon TLR challenge, reflecting loss of negative feedback control. This phenotype models dysregulated inflammatory responses relevant to sepsis, autoimmune disease, and metabolic disorders, making the cells a valuable tool for studying innate immune regulation.
These knockout cells facilitate detailed analysis of MAPK signaling dynamics and cytokine regulation. Applications include kinetic profiling of phospho-ERK1/2, phospho-JNK, and phospho-p38 by western blotting; cytokine mRNA and protein quantification via RT-qPCR and ELISA; immunofluorescence localization of DUSP1; and flow cytometric assessment of macrophage activation markers. The cells are also suited for AP-1/NF-??B luciferase reporter assays and high-throughput screening of kinase or phosphatase inhibitors. For further information, contact Ascent Research.
