APEX1 Knockout HEK293T Polyclonal Cells are a ready-to-use, CRISPR/Cas9-edited heterogeneous population of HEK293T cells featuring targeted disruption of the APEX1 gene. This polyclonal knockout cell pool serves as a versatile loss-of-function research model for investigating the multifunctional roles of the APEX1 protein in DNA repair and redox signaling, without the need for single-cell cloning or monoclonal characterization. The product is supplied as a viable, expanded cell population that can be immediately deployed in standard cell culture workflows, providing a convenient system for functional genomics, pathway dissection, and mechanistic studies.
HEK293T is a human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen, which promotes episomal replication of SV40 ori-containing plasmids and dramatically boosts transient protein expression. This feature, combined with its fast growth and high transfection efficiency, has made HEK293T a workhorse for lentivirus production, protein overexpression, and genome-scale genetic screens. The epithelial origin and embryonic kidney lineage also render it a relevant host for studying cellular responses to genotoxic and oxidative insults, as these pathways are broadly conserved across epithelial tissues.
APEX1 encodes a bifunctional protein that acts as an apurinic/apyrimidinic (AP) endonuclease in the base excision repair (BER) pathway, initiating the removal of abasic sites generated by spontaneous hydrolysis or oxidative damage, and as a redox cofactor that reduces critical cysteine residues on transcription factors to modulate their DNA-binding activity. Downstream of reactive oxygen species (ROS), APEX1 is transcriptionally regulated by TP53, HIF1A, and CREB1, and it physically interacts with BER partners XRCC1, POLB, and PCNA as well as with transcription factors RELA (NF-??B p65) and TP53. Through its redox function, APEX1 activates FOS/JUN (AP-1) and NFKB1, thereby coupling oxidative stress sensing to inflammatory and survival gene expression programs. This dual functionality positions APEX1 at a critical node linking genome maintenance with transcriptional control.
In the HEK293T background, disruption of APEX1 compromises both the repair of oxidative DNA lesions and the redox regulation of transcription factors, creating a sensitized cellular background for probing DNA damage response and redox biology. The polyclonal nature of the knockout pool mirrors the genetic heterogeneity inherent in many experimental setups, making it especially suitable for pooled functional assays and screens where clonal artifacts are undesirable. Moreover, the cell line’s high transfectability allows for convenient rescue experiments and ectopic expression of APEX1 variants, enabling structure-function studies directly in the knockout context.
This product can be applied across diverse research domains, including cancer biology, neurodegeneration, inflammation, and aging. Representative assays utilizing these cells include comet assay and BER activity measurements to quantify DNA repair capacity, redox activity assays, co-immunoprecipitation to map APEX1 interactions with XRCC1 or RELA, luciferase reporter assays for monitoring AP-1 and NF-??B transcriptional activity, and MTT-based viability tests under acute oxidative stress. The model is also ideal for CRISPR-based genetic interaction mapping and high-throughput drug resistance screening. For additional technical support, please contact Ascent Research.
