The EIF2AK2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the human Burkitt lymphoma Raji B lymphocyte line. This loss-of-function model targets EIF2AK2 (PKR), an interferon-inducible kinase pivotal to antiviral innate immunity and stress signaling. The heterogeneous knockout pool enables functional studies without clonal selection biases, providing a physiologically relevant system for investigating PKR-dependent pathways.
The Raji cell line is an EBV-positive, lymphoblastoid suspension culture derived from a Burkitt lymphoma patient. Widely used as a model for B cell malignancies and immune responses, Raji cells maintain intact interferon and NF-??B signaling cascades. These features render the cell line suitable for examining molecular interactions between viral sensing mechanisms and lymphomagenesis.
EIF2AK2 encodes a serine/threonine kinase activated by double-stranded RNA (dsRNA) during viral infection. Upon activation, PKR phosphorylates eIF2?? (EIF2S1) on Ser51, inhibiting global translation while promoting selective translation of stress-responsive mRNAs such as ATF4 and CHOP/DDIT3. PKR also stimulates NF-??B signaling via I??B?? phosphorylation and IKK activation, and promotes apoptosis through JNK/MAPK8 and p53/TP53 pathways. Regulatory inputs include interferons (IFNA, IFNB), viral dsRNA, PACT/PRKRA, and TLR3/7/8 agonists like poly(I:C). Key interacting partners encompass PACT, TRBP/TARBP2, p58IPK/DNAJC3, and STAT1, with downstream effectors including eIF2??, NF-??B, JNK, and p53. PKR operates within innate immune networks, linking RIG-I/MDA5-MAVS-TBK1-IKK?? sensing to interferon signaling via IFNAR1/2, JAK1, TYK2, STAT1/2, and IRF9.
In Raji B cells, EIF2AK2 disruption enables dissection of PKR??s dual roles in antiviral defense and tumor biology. As Raji cells harbor latent EBV, which produces dsRNA intermediates, this knockout model aids in studying PKR??s impact on viral latency and B cell transformation. It also facilitates investigation of the crosstalk between innate immunity and NF-??B-driven survival pathways in lymphoma.
These polyclonal knockout cells are suited for functional assays including Western blot analysis of phospho-eIF2?? (Ser51), RT-qPCR quantification of interferon-stimulated genes (ISGs), and luciferase-based NF-??B reporter assays. Additional methodologies include flow cytometry for apoptosis (Annexin V/PI), RNA-seq to assess translational efficiency, co-immunoprecipitation of PKR with dsRNA or PACT, viral replication assays (plaque assay, TCID50), and stress granule imaging. This model supports antiviral research, cancer biology, and drug screening efforts targeting the PKR pathway. For further information, please contact Ascent Research.
