The INIP Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited population of human liver adenocarcinoma cells bearing targeted disruption of the INIP gene. As a polyclonal knockout model, the product comprises a heterogeneous pool of INIP-deficient SK-HEP-1 derivatives generated by CRISPR-mediated gene disruption, avoiding single-cell cloning. This format enables robust analysis of INIP loss-of-function effects across a cell population, minimizing clonal biases in assays relevant to DNA damage signaling and cancer biology.
The parental SK-HEP-1 cell line was originally isolated from the ascitic fluid of a liver adenocarcinoma patient and displays an endothelial-like phenotype. It serves as a well-characterized model for hepatic tumor biology, including studies of cancer cell proliferation, metastasis, and therapeutic response. The liver adenocarcinoma background is particularly relevant for investigating DNA repair pathways in hepatocellular carcinoma, as these tumors frequently exhibit genomic instability and altered damage responses.
INIP is an essential component of the sensor of single-stranded DNA (SOSS) complex, where it interacts with INTS3 and NABP1 to recognize ssDNA at double-strand breaks. This complex stabilizes the recruited MRN complex (MRE11-RAD50-NBS1) and promotes ATM kinase activation. INIP-dependent ATM signaling phosphorylates downstream targets such as CHEK2, TP53, and H2AX, thereby enforcing the G2/M checkpoint and facilitating homologous recombination repair. Disruption of INIP destabilizes the SOSS complex, attenuates ATM pathway activation, and sensitizes cells to ionizing radiation and genotoxic agents.
In the SK-HEP-1 liver adenocarcinoma context, INIP knockout provides a powerful system to examine how DNA damage response defects influence hepatic cancer phenotypes. The polyclonal knockout population permits assessment of average cellular behaviors such as proliferation, survival, and migration under endogenous or therapy-induced genotoxic stress. This model facilitates the study of synthetic lethal interactions, drug resistance mechanisms, and the role of genome maintenance in liver cancer progression, without the confounding effects of clonal selection.
Key applications include DNA repair pathway dissection, synthetic lethality screening, and evaluation of chemotherapeutic sensitivity. The cells are suited for western blotting of phospho-ATM and ??-H2AX, immunofluorescence detection of ??-H2AX foci, clonogenic survival assays after irradiation, cell cycle flow cytometry, and comet assays. Co-immunoprecipitation can confirm loss of SOSS complex integrity by probing INTS3 and NABP1 interactions. Additionally, the model supports liver cancer metastasis studies via migration and invasion assays. Together, these INIP knockout cells serve as a versatile tool for both basic and translational oncology research. For further information, please contact Ascent Research.
