The EIF3C Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the EIF3C gene in the HEK293T human embryonic kidney cell line. This polyclonal population offers a heterogeneous loss-of-function model suitable for studying the roles of EIF3C in translation initiation and associated cellular processes without the need for single-cell cloning. The product provides a robust tool for functional genomics and pathway analysis, enabling researchers to interrogate the consequences of EIF3C disruption in a widely used cellular background.
HEK293T cells, originally derived from human embryonic kidney tissue, are transformed with adenovirus 5 DNA and stably express the SV40 large T antigen, which enhances episomal replication of plasmids containing the SV40 origin of replication. This cell line is extensively employed for recombinant protein expression, lentivirus production, and transient transfection studies due to its high transfectability and protein output. The kidney epithelial origin makes these cells relevant for investigations into renal biology and homeostasis, while their transformed nature provides a foundation for cancer and signaling research.
EIF3C encodes a core subunit of the eukaryotic translation initiation factor 3 (eIF3) complex, essential for cap-dependent translation initiation. It interacts with other eIF3 subunits, the 40S ribosomal subunit, and eIF4G to bridge the 43S pre-initiation complex with mRNA. Upstream regulators include mTORC1, MAPK/ERK, MYC, and signals such as amino acid availability and hypoxia. Downstream, EIF3C influences translation of Cyclin D1, c-MYC, and BCL2, linking translation control to cell cycle progression, proliferation, and apoptosis.
Disruption of EIF3C in HEK293T cells perturbs global protein synthesis and translation of mRNAs involved in cell cycle regulation and survival. The high proliferative capacity and transformed phenotype of HEK293T cells make this model ideal for studying translational reprogramming in oncogenic signaling and drug sensitivity. The polyclonal population provides a diverse loss-of-function background, valuable for pathway dissection and phenotypic screening in a kidney-derived epithelial context relevant to renal, breast, lung, colon, and gastric cancers.
This product supports a variety of experimental approaches, including polysome profiling to assess ribosome occupancy, puromycin incorporation assays to measure de novo protein synthesis, and co-immunoprecipitation to map eIF3 complex assembly. RT-qPCR and Western blotting enable validation of target gene expression and signaling changes, while cell proliferation and apoptosis assays facilitate functional characterization. Combined with RNA-seq for transcriptome-wide analysis, these tools aid investigations into translation regulation, mTOR signaling, cancer biology, and drug target validation. For additional information, please contact Ascent Research.
