The EIF4G3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of HAP1 cells, designed for loss-of-function studies of the human EIF4G3 gene. This product provides a heterogeneous pool of cells carrying gene disruptions, enabling bulk functional assays without the need for clonal isolation. The polyclonal format is convenient for population-level analyses of translation and signaling.
The HAP1 host cell line is a near-haploid, fibroblast-like cell line originally derived from the KBM-7 chronic myeloid leukemia (CML) cell line. They are adherent and possess a single copy of most chromosomes, which simplifies genetic knockout studies and enhances the efficiency of CRISPR/Cas9-mediated gene disruption. The CML origin provides a disease-relevant context for investigating cancer signaling pathways, making this model especially suitable for oncology research.
EIF4G3 encodes a large scaffold protein essential for assembly of the eIF4F translation initiation complex. It directly binds the cap-binding protein eIF4E, the RNA helicase eIF4A, and the multisubunit eIF3, thereby facilitating ribosome recruitment to the 5?? cap of mRNAs. EIF4G3 activity is regulated by the mTOR pathway through phosphorylation of 4E-BP1, which controls eIF4E availability. Additionally, EIF4G3 is phosphorylated by MNK1 downstream of MAPK signaling and interacts with poly(A)-binding protein (PABP) to mediate mRNA circularization. CRISPR/Cas9-mediated disruption of EIF4G3 impairs eIF4F complex formation, resulting in broad inhibition of cap-dependent translation initiation and altered expression of proteins linked to cell proliferation and stress responses.
In HAP1 cells, knockout of EIF4G3 provides a unique system to delineate the specific functions of this eIF4G paralog, as mammals encode three family members (EIF4G1, EIF4G2, and EIF4G3). The haploid genome ensures that disruption of the single EIF4G3 copy produces a loss-of-function phenotype, facilitating clean interpretation of knockout effects. This model is particularly valuable for studying translation control mechanisms in cancer, given the CML background, which offers a relevant platform for probing mTOR-driven oncogenic translation and identifying potential therapeutic targets in leukemia.
This polyclonal knockout population is suited to a range of experimental applications. Western blotting can confirm depletion of EIF4G3 and monitor associated translation factors such as eIF4E and eIF4A. Polysome profiling and puromycin incorporation assays directly assess global protein synthesis. RNA-sequencing coupled with ribosome profiling enables transcriptome-wide evaluation of translation efficiency. Cell viability and proliferation assays can measure functional consequences on growth and drug sensitivity. Collectively, these approaches support functional genomics of translation initiation, cancer drug target validation, and mechanistic dissection of mTOR signaling. For further details or technical assistance, please contact Ascent Research.
