This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from Raji cells, offering a heterogeneous pool of PDK1-disrupted genotypes (Homo sapiens). The polyclonal format maintains the inherent diversity of targeted gene disruption without clonal selection, enabling robust loss-of-function studies. Suitable for cancer and immunology research, these polyclonal knockout cells provide a flexible platform for investigating PDK1-dependent signaling in B cell malignancies and related contexts.
Raji cells are an EBV-positive B lymphocyte line established from a Burkitt lymphoma patient, extensively used as a model for B cell malignancies. The cells express surface IgM and B cell markers, retaining key B cell receptor (BCR) signaling machinery. Their transformed phenotype and well-characterized signaling pathways make them a valuable host for interrogating oncogenic and immune signaling networks, particularly in studies of lymphoma biology and therapeutic resistance.
PDK1 encodes a serine/threonine kinase that functions as a central node in the PI3K/AKT/mTOR signaling axis. Upon PI3K-mediated PIP3 production, PDK1 is recruited to the plasma membrane where it phosphorylates AKT at Thr308, a critical step for AKT activation. It also activates S6K (RPS6KB1) and RSK (RPS6KA), linking growth factor and BCR signaling to protein synthesis and cell proliferation. PDK1 interacts with HSP90 and 14-3-3 proteins, and is regulated by upstream signals from receptor tyrosine kinases, cytokine receptors, and the BCR via SYK. Downstream, PDK1-mediated AKT activation leads to mTORC1/S6K activation, NF-??B signaling, and FOXO transcription factor inhibition, collectively promoting cell survival, growth, and metabolic reprogramming.
In Raji B lymphocytes, PDK1 integrates BCR-derived signals to sustain the malignant phenotype. Knockout of PDK1 disrupts this signaling conduit, attenuating AKT phosphorylation and blunting downstream effectors such as S6K and NF-??B. Consequently, PDK1-deficient Raji cells exhibit reduced proliferation, increased susceptibility to apoptosis, and impaired metabolic adaptation. This model is thus invaluable for dissecting PDK1’s role in B cell lymphoma pathogenesis and for evaluating the dependency of lymphoma survival on PI3K/AKT pathway integrity.
Researchers can employ these cells in a wide range of assays, including Western blotting for pAKT-Thr308, flow cytometry to measure BCR-induced AKT activation, and MTT viability assays to assess drug responses. RT-qPCR profiling of downstream targets such as MYC and CCND1, Annexin V apoptosis staining, and co-immunoprecipitation of PDK1-AKT complexes further enable detailed mechanistic studies. The model is particularly suited for PI3K/AKT pathway inhibitor screening, BCR signal transduction analysis, and validation of lymphoma drug targets, and it facilitates cancer metabolism research and drug sensitivity testing. For additional information or to discuss custom applications, please contact Ascent Research.
