The CLK1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte line, harboring heterogeneous CLK1 gene disruptions. This format provides a pool of cells with diverse loss-of-function mutations, enabling functional studies of CLK1-dependent pre-mRNA splicing without clonal bias. The knockout abrogates CLK1 kinase activity, allowing researchers to dissect its role in alternative splicing, cell cycle progression, and apoptosis in a B-cell lymphoma context.
The parental Raji cell line is an EBV-positive Burkitt lymphoma line from a Nigerian male patient, exhibiting a mature B cell phenotype. It serves as a classic model for BCR signaling, lymphomagenesis, and apoptosis, with added relevance for studying EBV-driven oncogenesis. In this knockout, the Raji background provides a disease-appropriate system to evaluate CLK1??s contributions to splicing-mediated lymphoma survival.
CLK1 (CDC-like kinase 1) is a dual-specificity kinase that phosphorylates SR proteins, such as SRSF1, SRSF3, and SRSF4, regulating their function in pre-mRNA splicing within the spliceosome alongside U1 snRNP and U2AF. Its activity is controlled by upstream signals including MYC, the PI3K/AKT pathway, and cell cycle kinases. CLK1 interacts with SRSF proteins, PRPF4B, and SRPK1 to coordinate alternative splicing of apoptosis regulators (e.g., BCL2L1, MCL1) and metabolic enzymes (e.g., PKM). In Raji cells, loss of CLK1 disrupts this network, causing aberrant splicing of key apoptosis and cell cycle genes.
The CLK1 knockout in Raji cells models splicing dysregulation seen in B-cell lymphomas, where splicing factor alterations are common. This system enables investigation of how CLK1-dependent splicing events affect proliferation and apoptosis in a lymphoma-relevant background. The EBV-positive status further permits exploration of viral latency?Csplicing interplay, potentially uncovering novel therapeutic vulnerabilities.
Applications include screening CLK inhibitors, RNA-seq?Cbased splicing analysis, RT-qPCR of splice variants (e.g., BCL2L1, MCL1), phospho-SR protein western blotting, co-immunoprecipitation of CLK1 complexes, and functional assays such as Annexin V apoptosis staining, MTT proliferation, and cell cycle flow cytometry. This model also supports synthetic lethality studies with spliceosome inhibitors. For further information, contact Ascent Research.
