The EIF4A2 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of the AGS human gastric epithelial cell line, designed for functional disruption of the EIF4A2 gene. This heterogeneous pool of edited cells provides a physiologically relevant loss-of-function model for studying EIF4A2 in gastric cancer biology without selecting for a specific clonal genotype.
The AGS cell line is derived from a human gastric adenocarcinoma and is widely used as an in vitro model for gastric carcinoma. As adherent epithelial cells, AGS retains hallmark signaling aberrations, including altered mTOR pathway activity and dysregulated cap-dependent translation, making it a suitable background for analyzing RNA helicase-dependent gene expression.
EIF4A2 encodes an ATP-dependent DEAD-box RNA helicase that functions within the eIF4F complex, unwinding secondary structures in mRNA 5?? UTRs to facilitate ribosome loading and translation initiation. Its activity is inhibited by PDCD4 and promoted by mTORC1 signaling through 4E-BP1 phosphorylation and eIF4E activation. EIF4A2 interacts with eIF4G, eIF4B, and eIF4H to selectively enhance translation of mRNAs with structured 5?? UTRs, including those encoding CCND1, MYC, BCL2, and Survivin (BIRC5). Consequently, EIF4A2 disruption impairs the expression of these oncogenic factors, attenuating proliferative and survival signals.
In the AGS gastric cancer model, EIF4A2 knockout mimics pharmacological eIF4A inhibition, selectively reducing translation of oncogenic mRNAs while sparing those with unstructured 5?? UTRs. This polyclonal knockout population captures heterogeneous editing outcomes, enabling studies of gene function that reflect population-level cellular variability. The model allows dissection of translation control mechanisms unique to gastric adenocarcinoma and serves as a platform for evaluating target dependency and drug sensitivity.
Typical applications include proliferation and viability assays (MTT, colony formation), apoptosis detection (Annexin V), and immunoblotting for downstream targets CCND1, MYC, BCL2, and Survivin. Transcriptional and translational analyses by RT-qPCR and polysome profiling, as well as cap-binding affinity measurements, can be employed to dissect mechanistic consequences of EIF4A2 loss. The product is also suitable for eIF4A inhibitor screening, oncogene addiction studies, and mTOR pathway research. For further information or technical support, please contact Ascent Research.
