The DUSP14 Knockout HEK293T Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely utilized HEK293T human embryonic kidney cell line. These polyclonal knockout cells harbor targeted disruption of the DUSP14 gene, enabling loss-of-function studies of the dual-specificity phosphatase 14. The polyclonal format encompasses a heterogeneous mixture of edited alleles, reflecting the genetic diversity produced by CRISPR/Cas9-mediated gene disruption without clonal selection. This model is intended for researchers investigating DUSP14-dependent signaling mechanisms and cellular responses.
The HEK293T host cell line is a derivative of the HEK293 line that stably expresses the SV40 large T-antigen, facilitating episomal replication of plasmids and supporting high-efficiency transfection and viral packaging. Originating from human embryonic kidney tissue, HEK293T cells are a cornerstone in biomedical research for protein expression, lentiviral production, and transient transfection assays. Their robust growth and ease of manipulation make them an ideal background for generating knockout populations to dissect signaling pathways and protein function in a well-characterized cellular context.
DUSP14 encodes a dual-specificity phosphatase that directly dephosphorylates and inactivates JNK and ERK kinases, acting as a critical negative regulator of MAPK signaling. It is activated by upstream stimuli including TNF-?? and oxidative stress, and interacts with JIP scaffold proteins. By targeting JNK1 and ERK2, DUSP14 attenuates phosphorylation of c-Jun and ATF2, key AP-1 transcription factors, thereby modulating gene expression for proliferation, survival, and inflammatory responses. Thus, DUSP14 functions at the intersection of MAPK/ERK and JNK pathways, impacting T cell receptor signaling and apoptosis regulation.
HEK293T cells, widely used to study signal transduction, provide a robust background for assessing the impact of DUSP14 loss on MAPK pathway dynamics. This knockout population enables measurement of altered JNK and ERK activity, AP-1 transcriptional responses, and downstream effects on cell cycle and apoptosis. Such studies are pertinent to cancer, where MAPK dysregulation drives malignancy, and to autoimmune and inflammatory diseases, where aberrant signaling contributes to pathogenesis. The polyclonal format captures population-level responses, minimizing clonal variation artifacts.
Researchers can utilize these polyclonal knockout cells in diverse assays, including western blotting for DUSP14 and phospho-JNK/ERK after TNF-?? stimulation, RT-qPCR for AP-1 target genes, and AP-1 luciferase reporter assays. Phenotypic analyses like MTT proliferation, Annexin V apoptosis, and flow cytometric cell cycle assays can define DUSP14’s role in growth control. The model further supports drug resistance studies by evaluating chemotherapeutic sensitivity. For further information, please contact Ascent Research.
