APEX1 Knockout Huh-7 Polyclonal Cells are a precisely CRISPR/Cas9-edited polyclonal knockout population generated from Huh-7 human hepatocellular carcinoma cells, with targeted disruption of the APEX1 gene. This polyclonal pool facilitates loss-of-function studies of the dual-role APEX1 protein in base excision repair and redox signaling, without clonal selection bias.
The Huh-7 parental line, established from a liver tumor of a 57-year-old Japanese male in 1982, is a widely used and well-characterized human hepatocellular carcinoma model with epithelial morphology. Known for its relevance in liver cancer research, Huh-7 enables investigation of oncogenic pathways, drug metabolism, and therapeutic resistance in a hepatocyte-derived context.
APEX1 encodes a bifunctional enzyme: its abasic endonuclease domain cleaves the phosphodiester backbone 5?? to AP sites generated during BER, while the Ref-1 domain reduces oxidized cysteine residues in transcription factors, restoring DNA-binding activity. Key BER interactors include XRCC1, DNA polymerase beta, PARP1, PCNA, and DNA ligase III. Redox targets include AP-1 (Fos/Jun), NF-kB, p53, and HIF-1??. APEX1 is activated by oxidative stress, DNA damage, and kinases such as CK2 and PKC, and is transcriptionally regulated by Sp1 and p53, establishing it as a central integrator of DNA repair and stress-responsive transcriptional programs.
In hepatocellular carcinoma, APEX1 critically contributes to tumor maintenance, therapy resistance, and the oxidative stress response. Disruption of APEX1 in Huh-7 cells specifically allows interrogation of its role in BER proficiency, sensitization to DNA-damaging therapies, and redox-dependent transcription factor activation. This model is particularly valuable for testing BER inhibitors and elucidating links between chronic DNA damage, inflammation, and liver tumor biology.
Common downstream applications include APEX1 knockout confirmation via Western blot or RT-qPCR, DNA repair assays (alkaline comet, host cell reactivation), and oxidative stress sensitivity testing with H?O? or menadione. AP-1/NF-kB luciferase reporters assess transcriptional activity, while clonogenic survival and apoptosis assays evaluate responses to genotoxic agents. Drug sensitivity profiling with BER inhibitors such as methoxyamine or CRT0044876 supports translational research. For further inquiries, please contact Ascent Research.
