The GTF2H5 Knockout SK-HEP-1 Polyclonal Cells are a heterogeneous CRISPR/Cas9-edited population of human hepatic adenocarcinoma cells bearing targeted disruption of the GTF2H5 gene. This polyclonal knockout pool comprises a mixture of edited cell variants derived from the SK-HEP-1 host line, offering a robust loss-of-function model for studying the p8/TTDA protein without clonal selection bias. The format ensures representation of diverse genetic edits, making it suitable for functional studies in DNA repair and transcription.
SK-HEP-1 is a human liver adenocarcinoma cell line isolated from ascitic fluid of a patient with hepatic malignancy, and it displays both epithelial and endothelial-like characteristics. Widely adopted in hepatocellular carcinoma research, this adherent line provides a well-characterized cancer background for drug sensitivity screening, DNA damage response assays, and transcriptomic analyses. Its established molecular profile facilitates integration into standard cell-based experimental workflows.
GTF2H5 encodes p8/TTDA, a small stabilizing subunit of the transcription factor IIH (TFIIH) complex that is essential for RNA polymerase II transcription initiation and nucleotide excision repair (NER). p8 participates in both global genome and transcription-coupled NER subpathways. Upstream, its function is modulated by DNA damage signals including UV photoproducts and chemical adducts, via CDK7-mediated phosphorylation and ATR/ATM signaling. Within TFIIH, p8 interacts with core subunits such as XPB, XPD, p62, CDK7, and Cyclin H, and cooperates with XPA and RPA at damage sites. Downstream, functional TFIIH drives RNA polymerase II transcription, catalyzes repair synthesis, and contributes to p53 stabilization upon genotoxic stress.
Disruption of GTF2H5 in SK-HEP-1 liver cancer cells cripples NER capacity, leading to heightened sensitivity to ultraviolet radiation and DNA-damaging chemotherapeutics, and promotes genomic instability. This polyclonal knockout model is therefore valuable for dissecting NER deficiency in hepatic malignancy, exploring synthetic lethal vulnerabilities, and assessing how compromised transcription-coupled repair affects cancer cell proliferation and survival.
This GTF2H5 knockout cell pool empowers diverse investigations, including screening DNA-damaging agents for context-dependent toxicity, studying the molecular coupling between transcription and repair, and modeling trichothiodystrophy-associated cellular phenotypes. Researchers can validate knockout efficiency by western blotting and RT-qPCR, quantify NER activity via UV sensitivity and host cell reactivation assays, and monitor DNA damage accumulation using ??H2AX immunofluorescence and comet assays. Comprehensive transcriptome profiling by RNA-seq and flow cytometry-based cell cycle and apoptosis analyses further deepen mechanistic insights. For additional information or to place an order, please contact Ascent Research.
