The IER5 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in the HEK293T background, providing a heterogeneous pool of cells with targeted disruption of the IER5 gene. This loss-of-function model enables functional studies without single-cell cloning, facilitating efficient genotype-phenotype analysis in a population context. The knockout abolishes IER5 protein expression, allowing researchers to probe IER5-dependent molecular mechanisms.
HEK293T cells are human embryonic kidney epithelial derivatives stably expressing the SV40 large T antigen, which enhances episomal replication and supports high-yield protein expression, viral packaging, and transient transfection. This well-characterized line grows robustly and transfects with high efficiency, offering a reliable platform for gene function studies. While HEK293T cells retain functional p53, the large T antigen partially inactivates p53 and Rb pathways, creating a unique context for investigating stress-responsive genes like IER5.
IER5 is a p53-inducible transcriptional regulator and PP2A regulatory subunit. Upon DNA damage, ATM/ATR signal through p53 to induce IER5 expression. IER5 then interacts with the PP2A catalytic subunit (PPP2CA) and B55 regulatory subunit (PPP2R2A) to dephosphorylate c-Myc, promoting its degradation. This suppresses cell cycle progression and facilitates apoptosis, linking stress signaling to PP2A substrate selectivity.
In HEK293T cells, where SV40 large T antigen partially compromises p53, IER5 knockout allows dissection of p53-dependent and -independent roles in stress responses. Loss of IER5 may alter PP2A activity, stabilize c-Myc, and affect cell cycle checkpoints. This polyclonal population captures heterogeneous knockout effects, useful for studying dominant phenotypes, compensatory pathways, and population-level responses to DNA-damaging agents. The model is well-suited for examining radiation sensitivity and chemotherapeutic drug action in a genetically uniform yet functionally diverse system.
Applications include investigation of p53-mediated stress responses, PP2A substrate dephosphorylation, and c-Myc regulation in cancer biology. Compatible assays: Western blotting and RT-qPCR for gene/protein expression; co-immunoprecipitation to probe PP2A interactions; phospho-c-Myc analysis to assess PP2A activity; Annexin V apoptosis and cell cycle flow cytometry; and clonogenic survival for radiation/drug sensitivity. These cells also support drug discovery screens targeting PP2A or c-Myc pathways. For further details, contact Ascent Research.
