The EIF4A2 knockout Raji polyclonal cells provide a CRISPR/Cas9-edited polyclonal knockout cell population for targeted disruption of the EIF4A2 gene in the Raji B lymphocyte background. The polyclonal format ensures a heterogeneous loss-of-function model, preserving population-level diversity while avoiding clonal artifacts inherent to single-cell-derived knockout lines. This reagent is designed for functional genomic studies in pooled assays where biological variability is advantageous.
The Raji cell line is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma model that expresses CD19 and is extensively employed in B cell biology and cancer research. Derived from a pediatric patient, these cells retain mature B lymphocyte characteristics including antibody production and antigen presentation, while exhibiting aggressive proliferation driven by dysregulated oncogenic pathways typical of lymphomas.
EIF4A2 encodes the catalytic ATP-dependent RNA helicase subunit of the eIF4F translation initiation complex. Within this complex, EIF4A2 interacts with the cap-binding protein eIF4E and the scaffolding protein eIF4G to unwind structured 5?? UTRs of mRNAs, enabling ribosome scanning and cap-dependent translation. Its activity is positively regulated by mTORC1 signaling through 4E-BP1 phosphorylation and eIF4E release, as well as by MNK kinases downstream of growth factors such as EGF and insulin. The tumor suppressor PDCD4 negatively regulates EIF4A2 via direct binding. Key downstream targets include oncogenic mRNAs with complex 5?? UTRs, notably MYC, BCL2, and CCND1. Additional interacting proteins??eIF4B, eIF4H, and PABPC1??enhance helicase activity and translation initiation efficiency.
In the Raji B-cell lymphoma context, EIF4A2-mediated translational control is critical for the expression of survival and proliferation factors. Hyperactive mTOR signaling, a common feature in lymphomas, likely drives EIF4A2-dependent translation of MYC and BCL2, contributing to oncogenic growth. Disruption of EIF4A2 in this model permits dissection of cap-dependent translation initiation in B-cell malignancies and evaluation of the eIF4F complex??s role in lymphomagenesis. This polyclonal knockout population allows researchers to investigate how translation pathway components influence malignant B cell behavior.
Typical applications include polysome profiling to measure global translation, cap-dependent luciferase reporter assays, and flow cytometry for apoptosis and proliferation. The cells are compatible with RNA-seq and ribosome profiling for translatome analysis, co-immunoprecipitation and western blotting for protein interaction studies, and drug sensitivity testing with translation inhibitors like rocaglates and pateamine A. They can also be employed in CRISPR-based synthetic lethality screens to uncover targets that are selectively essential in EIF4A2-deficient lymphoma cells. For additional information, please contact Ascent Research.
