The DUSP23 Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the human DUSP23 gene has been disrupted to create a loss-of-function model. This product comprises a heterogeneous population of Jurkat T lymphocytes carrying a range of gene edits, providing a robust system for studying DUSP23-dependent signaling without the constraints of clonal selection. The polyclonal format preserves the cellular diversity inherent to the knockout process, enabling the investigation of DUSP23 function in a context that more closely reflects the natural variation of T-cell populations.
The parental Jurkat cell line is an immortalized human T-cell acute lymphoblastic leukemia line derived from a 14-year-old male. Jurkat cells express a functional TCR complex and are IL-2 dependent, serving as a standard model for TCR signaling, apoptosis, and leukemogenesis. Key proximal signaling molecules include ZAP-70, LAT, SLP-76, and PLC??1, which couple TCR activation to MAPK pathways such as ERK, JNK, and p38.
DUSP23 encodes a dual-specificity protein phosphatase that terminates JNK and p38 MAP kinase signaling by dephosphorylating phosphotyrosine and phosphothreonine residues. Activated downstream of TCR/CD3 stimulation and PMA/ionomycin, it interacts with JIP-1 (MAPK8IP1) and physically associates with JNK1, JNK2, and p38??. Through its phosphatase activity, DUSP23 inactivates these kinases, reducing phosphorylation of c-Jun, ATF2, and MAPKAPK2. This attenuates AP-1 transcriptional output and dampens T-cell activation, serving as a critical negative feedback node in the MAP kinase phosphatase network.
In Jurkat T cells, DUSP23 knockout removes a brake on JNK/p38 signaling, potentially leading to sustained phosphorylation of these kinases and enhanced activation of c-Jun, ATF2, and MAPKAPK2 following TCR stimulation. This may result in increased AP-1 activity and altered expression of activation markers like CD69 and CD25. Such dysregulation is relevant to T-cell acute lymphoblastic leukemia, where aberrant MAPK signaling drives proliferation, and to autoimmune diseases characterized by overactive TCR responses. The polyclonal population minimizes clonal artifacts, ensuring robust functional analysis.
Applications include mechanistic studies of JNK/p38 regulation via Western blotting for phospho-JNK and phospho-p38, RT-qPCR for c-Jun and ATF2, and flow cytometry for activation markers. The knockout cells support AP-1/NF-??B reporter assays, co-immunoprecipitation of JIP-1 interaction, and viability/apoptosis assays with pathway inhibitors. Cytokine secretion (IL-2) assays can reveal functional consequences of DUSP23 loss. This model is valuable for MAPK modulator screening in leukemia and autoimmunity. For inquiries, please contact Ascent Research.
