The EIF4A2 Knockout SK-HEP-1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of SK-HEP-1 cells harboring targeted disruption of the EIF4A2 gene. This polyclonal knockout model provides a heterogeneous pool of gene-edited cells suitable for studying loss-of-function effects without selection of a single clone. The resulting cell population offers a versatile tool for investigating EIF4A2-dependent processes in a human hepatic adenocarcinoma background.
The parental SK-HEP-1 cell line is an adherent epithelial cell model established from the liver adenocarcinoma of a 52-year-old male patient. These cells retain malignant characteristics of hepatic origin and are widely employed as an in vitro model for hepatocellular carcinoma research. Their defined genetic background and reproducible growth properties make them a robust platform for evaluating gene function in liver cancer biology.
EIF4A2 encodes an ATP-dependent DEAD-box RNA helicase that functions as a core component of the eIF4F translation initiation complex. Together with the cap-binding protein eIF4E and the scaffold eIF4G, EIF4A2 unwinds secondary structures within 5?? untranslated regions of mRNAs, facilitating 43S preinitiation complex recruitment and cap-dependent translation initiation. Its helicase activity is stimulated by eIF4B, eIF4H, and the co-activator DDX3. EIF4A2 expression and activity are regulated by growth factor signaling, notably through mTORC1-mediated phosphorylation of 4E-BP1 that releases eIF4E to assemble the eIF4F complex. Upstream signals include the PI3K/AKT and MAPK/ERK pathways, and transcription factors such as MYC and HIF1A, whereas downstream targets encompass a subset of proto-oncogenes with highly structured 5?? UTRs, including CCND1, MYC, BCL2, and VEGFA.
Disruption of EIF4A2 in the SK-HEP-1 hepatocellular carcinoma model creates a powerful system to dissect the contribution of cap-dependent translation to liver cancer cell proliferation, survival, and metastatic potential. The polyclonal knockout population enables assessment of heterogeneity in translation control and allows for bulk functional studies without clonal selection artifacts. This model is particularly relevant for investigating the dependency of liver cancer cells on eIF4F complex activity, which is often dysregulated in tumors with hyperactive mTOR signaling.
Researchers can use this knockout model to investigate cap-dependent translation mechanisms in liver cancer by polysome profiling and m7GTP cap-binding assays, or to screen translation inhibitors such as silvestrol and rocaglates in proliferation and drug sensitivity assays. The cells allow monitoring of mTOR signaling via phospho-4E-BP1 and phospho-S6 levels, and assessment of EIF4A2 in migration and invasion using transwell assays. In vivo xenografts evaluate tumor growth and metastasis upon EIF4A2 loss. RNA-seq and dual luciferase reporters assess global and targeted translation changes. For technical support, contact Ascent Research.
