The EIF4G3 Knockout HEK293T Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HEK293T cells with disrupted EIF4G3 gene function. This heterogeneous knockout pool serves as a loss-of-function model for interrogating translation initiation pathways. The polyclonal format provides a breadth of editing events to support robust, population-level analysis.
HEK293T cells are an adherent human embryonic kidney epithelial line transformed with the SV40 large T antigen, enabling episomal plasmid replication and high transfection efficiency. Their rapid growth and amenability to diverse biochemical and genetic assays have made them a workhorse for signal transduction and translation research.
EIF4G3 functions as a central scaffold within the eIF4F complex, bridging the cap-binding protein eIF4E, the RNA helicase eIF4A, and eIF3 to recruit the 43S preinitiation complex. This interaction drives cap-dependent translation initiation and facilitates mRNA circularization through binding of PABPC1. Under stress, EIF4G3 can support IRES-mediated translation of select mRNAs, including c-Myc, VEGF, and XIAP. Its activity is regulated by the mTORC1 kinase in response to growth factors, amino acids, and cellular energy status, with upstream inputs from AMPK and ER stress sensors converging on the 4E-BP/S6K axis. EIF4G3 also associates with MNK1/2 kinases that phosphorylate eIF4E, further modulating cap-dependent initiation efficiency.
Disruption of EIF4G3 in the HEK293T background permits direct dissection of cap-dependent versus IRES-dependent translation mechanisms in a tractable cell system. Because HEK293T cells exhibit robust basal translation, EIF4G3 loss reveals the dependency of specific transcripts on intact eIF4F assembly and allows assessment of mTOR pathway outputs under defined conditions. The polyclonal knockout nature provides a population-level readout, making it ideal for comparing translation phenotypes in the absence of a scaffold protein that integrates multiple signaling inputs.
This product is applicable to assays such as polysome profiling, cap-dependent dual luciferase reporter analysis, puromycin incorporation, and S35-methionine metabolic labeling. It also enables co-immunoprecipitation of eIF4F components to evaluate complex formation, RNA immunoprecipitation to identify target mRNAs, and inhibitor screens targeting translation initiation. For technical details or ordering, contact Ascent Research.
