The DUSP1 Knockout HEK293T Polyclonal Cells product offers a polyclonal population of CRISPR/Cas9-edited HEK293T cells with targeted disruption of the DUSP1 gene, providing a loss-of-function model for the MKP-1 phosphatase. This polyclonal knockout pool ensures robust attenuation of MKP-1 expression across the population without the need for single-cell cloning, delivering a flexible and reliable system for investigating MAPK signaling dynamics.
The host HEK293T cell line, derived from human embryonic kidney cells, stably expresses the SV40 large T antigen, which facilitates high-level plasmid replication and protein production. With its epithelial-like morphology and excellent transfectability, HEK293T is ideal for protein expression, viral packaging, and detailed signaling studies, forming a well-characterized background for gene-edited knockout models.
DUSP1 encodes MKP-1, a dual-specificity phosphatase that dephosphorylates threonine and tyrosine residues in the activation loop of MAP kinases, including ERK1/2 (MAPK1/3), JNK (MAPK8/9), and p38 (MAPK14), thereby attenuating their signaling. It functions downstream of diverse upstream stimuli such as EGF, TNF-??, oxidative stress, and transcription factors p53 and NF-??B. Through its phosphatase activity, MKP-1 regulates the phosphorylation status of downstream effectors like c-Jun, ATF2, and Elk-1 and impacts the expression of pro-inflammatory cytokines (e.g., IL-6) and cell cycle regulators. Within the MAPK cascade, RAS, RAF, and MEK act upstream of ERK, JNK, and p38, positioning DUSP1 as a key negative feedback regulator. Consequently, disruption of DUSP1 is expected to result in sustained MAPK activation, altering cellular responses to growth factors and stress.
In HEK293T cells, DUSP1 knockout leads to enhanced and prolonged MAPK activation upon stimulation, altering proliferation, differentiation, and cytokine responses. This provides a model to study feedback mechanisms and signaling thresholds. High transfectability enables co-expression experiments for rescue or domain mapping. The absence of MKP-1 offers a clean background to examine interactions with MAPKAPKs and cross-talk with other pathways.
Researchers can use these polyclonal knockout cells for Western blotting of phospho-ERK, -p38, and -JNK, RT-qPCR analyses of DUSP1 and targets like c-Fos and IL-6, immunofluorescence for MAPK localization, and proliferation or cytokine assays. Applications include phospho-signaling studies, drug screening for MAPK inhibitors, and functional genomics in cancer, inflammation, metabolic, or neurodegenerative research. Contact Ascent Research for details.
