The CDK8 Knockout Raji Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function investigation of CDK8 in a human B lymphocyte background. This suspension-adapted pool contains a heterogeneous array of edited alleles arising from target-gene disruption, delivering a genetically diverse knockout model without single-cell cloning. Supplied as stable, proliferating Raji cells, the population allows robust biochemical and functional interrogation of CDK8-dependent processes within a well-characterized Burkitt lymphoma-derived system.
The host cell line, Raji, is an EBV-positive B lymphocyte line established from a Burkitt lymphoma patient. It exhibits suspension growth and endogenously expresses the B cell surface markers CD19 and CD20, rendering it an established model for B cell receptor signaling and lymphomagenesis. The EBV-positive status recapitulates aspects of viral-driven B cell transformation, providing a disease-relevant context for dissecting oncogenic networks and the interplay between host transcription and viral latency programs.
CDK8 functions as the catalytic kinase subunit of the CDK8 module within the Mediator complex, where it phosphorylates the C-terminal domain of RNA polymerase II and transcription factors, including STAT1 and SMAD2/3, to modulate gene expression. Upstream signals from Wnt ligands, TGF-??1, and EGF converge through ??-catenin, SMADs, and STAT1, placing CDK8 at a signaling nexus that drives transcription of MYC, CCND1, and FOSL1 while repressing CDKN1A and BAX. Key interacting partners Cyclin C, MED12, MED13, and E2F1 fine-tune Mediator recruitment, so CDK8 knockout uncouples the complex from Wnt/??-catenin, TGF-??/SMAD, and p53 pathways, thereby altering transcriptional programs governing proliferation and apoptosis.
Within Burkitt lymphoma, CDK8 disruption is predicted to impair oncogenic transcription driven by dysregulated ??-catenin and MYC, potentially restoring tumor-suppressive signals mediated by p53 and TGF-??. This knockout resource enables direct interrogation of CDK8??s role in sustaining lymphoma cell growth and survival, and its interaction with EBV latency. Moreover, given CDK8??s involvement in colorectal cancer, melanoma, and acute myeloid leukemia, findings in this B cell model may offer broader mechanistic insights into CDK8-dependent transcription in cancer.
Typical applications include functional dissection of Mediator kinase activity in B cell malignancies, validation of CDK8-targeted inhibitors, and genome-wide transcriptomic analyses via RNA-seq or ChIP-qPCR to evaluate RNA polymerase II occupancy. Proliferation and apoptosis can be assessed through MTT and Annexin V assays, while phospho-STAT1 signaling is monitored by Western blot or flow cytometry. Co-immunoprecipitation of CDK8 with MED12 confirms module integrity, and RT-qPCR quantifies changes in MYC, CCND1, and CDKN1A expression. The polyclonal format is well-suited for pathway epistasis studies using Wnt3a or TGF-??1 stimulation and for high-throughput screening campaigns. For further information or custom requests, contact Ascent Research.
