The MAP3K12 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the MAP3K12 gene in a B-lymphocyte context. The polyclonal nature provides a heterogeneous pool of Raji cells with diverse gene disruptions, enabling robust interrogation of MAP3K12-dependent signaling without clonal selection artifacts. This format is ideal for genotype-phenotype correlation, functional genomics screens, and pathway dissection where population-level responses are paramount.
The Raji cell line is an EBV-positive B lymphoblastoid line derived from a Burkitt lymphoma patient. These suspension cells carry the t(8;14) translocation, leading to c-MYC dysregulation, and serve as a validated model for B-cell lymphoma biology, oncogenic signaling, and EBV latency. Raji cells are well-suited for functional genomic manipulation and biochemical assays, including high-throughput screening.
MAP3K12, also known as dual leucine zipper kinase (DLK), encodes a serine/threonine kinase that functions as an upstream activator of the stress-activated c-Jun N-terminal kinase (JNK) signaling module. Upon stimulation by upstream signals such as tumor necrosis factor-alpha (TNF-??), interleukin-1 (IL-1), or cellular stressors including ultraviolet radiation and oxidative stress, MAP3K12 directly phosphorylates and activates MAP2K7 (MKK7). Activated MKK7 subsequently phosphorylates JNK1 and JNK2, which in turn phosphorylate key transcription factors such as c-Jun and ATF2, promoting their transactivation activity. The scaffolding proteins JIP-1 and JIP-2 facilitate the assembly of this kinase cascade, while the adaptor protein TRAF2 links MAP3K12 to upstream cytokine receptors. This pathway ultimately regulates gene expression programs governing apoptosis, cell differentiation, and proliferation in response to stress and immune signals.
In Raji B-cell lymphoma, MAP3K12-dependent JNK signaling is critical for apoptosis, survival, and cytokine responses. Disruption of MAP3K12 allows dissection of DLK-JNK contributions to EBV-transformed B-cell biology, including stress-induced death pathways and c-MYC-driven proliferation. This polyclonal knockout population is a robust tool for studying MAP3K12 function in the context of constitutive NF-??B and latent viral gene expression typical of EBV-positive lymphomas, revealing potential therapeutic vulnerabilities.
This reagent supports a broad range of applications: investigation of JNK signaling dynamics in B-cell lymphoma, functional dissection of stress kinase pathways, and high-throughput inhibitor screening. Representative assays include phospho-JNK and phospho-c-Jun Western blotting, AP-1 luciferase reporter assay, Annexin V apoptosis assay, RT-qPCR for JNK target genes, and co-immunoprecipitation of MKK7. The polyclonal format enables efficient evaluation of MAP3K12-dependent phenotypes without clonal variation. For technical details, contact Ascent Research.
