EML4 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji human B-lymphocyte cell line, designed for the study of EML4 gene function through targeted gene disruption. This polyclonal population contains a heterogeneous mixture of knockout alleles, generated by CRISPR/Cas9-mediated cleavage without single-cell clonal expansion, providing a representative loss-of-function model that avoids clonal artifacts. The product enables investigation of endogenous EML4 biology in a B-cell lymphoma context, supporting applications in cancer cell biology, immunology, and drug discovery.
The Raji cell line is an EBV-positive lymphoblastoid line established from a Burkitt lymphoma patient, widely used as a model for B-cell malignancies and Epstein-Barr virus biology. These cells express canonical B-cell markers including CD19 and CD20, and maintain a mature B-cell phenotype. As a suspension cell line with robust proliferative capacity, Raji cells are amenable to scalable experimental workflows, including high-throughput screening, functional genomics, and biochemical assays that require consistent genetic backgrounds.
EML4 encodes a microtubule-associated protein that directly interacts with ??/??-tubulin heterodimers to regulate microtubule polymerization and dynamics. Its activity is tightly controlled during the cell cycle: CDK1 and Aurora A kinases phosphorylate EML4, modulating its microtubule-binding affinity and localization to the mitotic spindle. Acting downstream of these mitotic kinases, EML4 participates in spindle assembly, chromosome alignment, and mitotic progression by forming complexes with NEK6 and NEK7 kinases, which further influence microtubule stability. In certain oncogenic contexts, EML4 forms a fusion with ALK, generating a constitutively active tyrosine kinase that drives non-small cell lung cancer and other malignancies.
In the Raji B-cell context, EML4 knockout allows dissection of its contribution to lymphocyte-specific processes such as antigen receptor signaling, cell division, and genomic stability. Given the EBV-driven proliferative nature of Raji cells, loss of a microtubule regulator may impact cell cycle checkpoints and susceptibility to genomic instability, providing insights into lymphomagenesis mechanisms. Furthermore, this polyclonal knockout line serves as an ideal genetic background for reconstitution experiments, including expression of wild-type EML4 or the oncogenic EML4-ALK fusion, facilitating structure-function analyses and comparative studies in a human B-cell environment.
This product is suited for a range of experimental applications, including functional characterization of EML4 in lymphocyte biology through Western blotting and RT-qPCR confirmation of knockout, immunofluorescence microscopy to assess spindle morphology using ??-tubulin staining, and flow cytometry-based cell cycle analysis. It also supports investigation of microtubule-targeting agents and high-throughput drug screening with ALK inhibitors such as crizotinib, using proliferation (MTS) and apoptosis (Annexin V) assays. Drug sensitivity screens can be performed to evaluate synergistic or antagonistic interactions in B-cell malignancies. For additional technical details or support regarding this product, please contact Ascent Research.
