The EIF4A2 Knockout Jurkat Polyclonal Cells represent a genetically engineered Jurkat T-lymphocyte population carrying CRISPR/Cas9-mediated disruption of the EIF4A2 gene. This polyclonal knockout model provides a heterogeneous pool of edited cells, enabling loss-of-function studies of the EIF4A2 ATP-dependent RNA helicase in a human T-cell acute lymphoblastic leukemia (T-ALL) background. The product is supplied as viable, cryopreserved polyclonal knockout cells suitable for immediate culture and downstream functional analyses without subcloning requirements.
The parental Jurkat cell line is an immortalized human T-cell line established from the peripheral blood of a T-ALL patient. Jurkat cells are extensively used in immunological research, particularly for studying T-cell receptor (TCR) signaling pathways, activation mechanisms, and apoptosis. Their leukemic origin also makes them a relevant model for T-ALL and other hematological malignancies. These suspension cells maintain key features of T-cell biology, including expression of CD3 and other surface markers, and they remain susceptible to genetic manipulation, making them an ideal host for knockout generation.
EIF4A2 functions as an ATP-dependent RNA helicase and a core component of the eIF4F translation initiation complex, along with the cap-binding protein eIF4E and scaffold eIF4G1. It unwinds secondary structures in 5′ UTRs to facilitate ribosomal scanning, enabling efficient cap-dependent translation of oncogenic mRNAs such as MYC, CCND1, BCL2, and VEGFA. Its activity is tightly regulated by upstream mTORC1 kinase signaling via phosphorylation of 4E-BP1 and S6K1, and is further modulated by the PI3K/AKT and RAS/ERK/MNK1 cascades. Transcription factors MYC and HIF1A promote EIF4A2 expression, while PDCD4 acts as a direct inhibitor. Accessory factors eIF4B and eIF4H enhance helicase processivity.
In Jurkat T-ALL cells, EIF4A2 knockout disrupts cap-dependent translation of mRNAs with structured 5′ UTRs, potentially downregulating oncogenic drivers like MYC and cyclin D1, which are critical for leukemic proliferation and survival. This model enables dissection of how translational control through eIF4F intersects with TCR signaling and mTOR pathway activity. Additionally, EIF4A2 loss may sensitize cells to the translation inhibitor silvestrol, providing a tool for drug efficacy studies.
These polyclonal knockout cells are suited for a broad range of experimental applications. Translation regulation can be analyzed by polysome profiling, cap-binding assays, and 5′ UTR reporter systems. T-cell activation studies may employ flow cytometry to measure TCR-induced activation markers and cytokine output. Proliferation, apoptosis, and cell cycle assays (e.g., MTT, annexin V, EdU) assess leukemic growth properties. Western blotting and RT-qPCR can quantify MYC, CCND1, and BCL2 expression, while phospho-specific antibodies monitor mTOR pathway status. Drug sensitivity screening with silvestrol or mTOR inhibitors reveals translational dependencies. RNA-seq provides transcriptome-wide insights. For further information, please contact Ascent Research.
