The DUSP22 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human embryonic kidney HEK293T cell line, enabling loss-of-function studies of the dual-specificity phosphatase DUSP22. These polyclonal cells harbor a targeted disruption of the DUSP22 gene, resulting in ablation of endogenous DUSP22 protein and providing a robust model for dissecting DUSP22-dependent signaling without pharmacological artifacts. The polyclonal format maintains genetic diversity while ensuring consistent gene disruption across the pool, mitigating clonal variation.
HEK293T is a widely utilized mammalian cell line characterized by high transfection efficiency and robust growth, owed to stable expression of the SV40 large T antigen that permits episomal replication of SV40 origin?Ccontaining plasmids. Derived from human embryonic kidney cells transformed with sheared adenovirus 5 DNA, this epithelial line excels in recombinant protein expression, lentivirus production, and gene-editing applications, providing a versatile background for studying signaling pathways and cancer biology.
DUSP22 (JNK/SAPK-associated phosphatase) is a dual-specificity phosphatase that negatively regulates MAPK signaling by dephosphorylating JNK (MAPK8/9), p38?? (MAPK14), and ERK2 (MAPK1) at conserved activation-loop residues. In TCR signaling, DUSP22 is recruited to the immunological synapse, where it dephosphorylates the kinase Lck to attenuate proximal signaling. DUSP22 also modulates STAT3 activity and interacts with ASK1 (MAP3K5). Upstream, DUSP22 is controlled by TCR activation, CD28 co-stimulation, oxidative stress, and AP-1/NFAT transcription factors; downstream, it influences AP-1-mediated transcription and NF-??B via I??B?? (NFKBIA).
In the HEK293T background, which lacks endogenous T-cell machinery, the DUSP22 knockout model offers a clean platform for studying core MAPK and STAT3 regulatory mechanisms. The high transfectability of HEK293T facilitates reconstitution of DUSP22 variants or pathway components for structure-function analyses and interaction mapping with partners like JNK, p38, ERK, and STAT3. This system is particularly relevant for exploring DUSP22 dysregulation in lymphomas, autoimmune diseases, and inflammation.
Applications include biochemical analyses such as Western blotting for phospho-MAPKs, co-immunoprecipitation of DUSP22 with MAPK substrates, and dual-luciferase reporter assays for AP-1/NF-??B activity. Transcriptional profiling by RT-qPCR for FOS and JUN and flow cytometry for phospho-STAT3 complement these studies. The cells support functional complementation with mutants, substrate screening, and validation of DUSP22 as a therapeutic target in cancer and immune disorders. For further information or technical support, please contact Ascent Research.
