The ITPA Knockout SK-HEP-1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population in which the ITPA gene has been disrupted. This model is generated on the SK-HEP-1 human hepatic adenocarcinoma cell line and serves as a valuable tool for investigating the roles of inosine triphosphate pyrophosphatase in nucleotide metabolism, thiopurine drug response, and maintenance of genomic stability. The polyclonal population reflects a heterogeneous mix of cells bearing diverse CRISPR-mediated edits, enabling the study of ITPA loss-of-function in a context that retains the genetic diversity of an uncloned pool.
SK-HEP-1 cells were originally derived from the ascitic fluid of a patient with liver adenocarcinoma and have been widely employed as a model for hepatocellular carcinoma research. Notable for their endothelial-like characteristics, these cells exhibit a unique phenotype that renders them suitable for studies of liver cancer biology, metastatic behavior, and angiogenesis. The host cell line??s well-characterized growth properties and compatibility with standard culture conditions make it an accessible platform for functional genomics and pharmacological screening.
ITPA encodes inosine triphosphate pyrophosphatase, an enzyme that hydrolyzes inosine triphosphate (ITP) and deoxyinosine triphosphate (dITP) to their corresponding monophosphates, thereby preventing the accumulation and misincorporation of noncanonical nucleotides into DNA and RNA. Within the purine salvage pathway, ITPA functions downstream of nucleotide diphosphate kinases and interacts with key metabolic enzymes such as inosine-5??-monophosphate dehydrogenase (IMPDH) and hypoxanthine-guanine phosphoribosyltransferase 1 (HPRT1). The enzyme??s activity is modulated by oxidative stress through NRF2-mediated transcriptional regulation, and its loss leads to elevated intracellular ITP/dITP levels, increased formation of aberrant nucleotides, and heightened sensitivity to thiopurine prodrugs such as azathioprine and 6-mercaptopurine. Consequently, ITPA deficiency is associated with thiopurine-induced toxicity and has been implicated in certain neurodevelopmental disorders.
Disruption of ITPA in the SK-HEP-1 hepatic cancer background creates a physiologically relevant model for exploring the intersection between purine nucleotide imbalance, genomic instability, and liver tumor biology. The knockout cells allow researchers to dissect how accumulated ITP affects DNA replication fidelity, base excision repair, and the response to oxidative DNA damage within a cancer cell environment. Because hepatic adenocarcinoma cells exhibit distinct metabolic adaptations, this model is particularly suited for studying nucleotide pool perturbations and their impact on cancer cell proliferation, drug resistance, and redox homeostasis.
This ITPA knockout cell pool is applicable to a wide range of experimental approaches, including thiopurine pharmacogenomics, screening of purine metabolism modulators, and investigation of oxidative stress-mediated signaling. Researchers can assess ITPA protein expression via western blotting, quantify ITP/IMP levels by LC-MS, evaluate DNA damage through comet assays or ??H2AX immunofluorescence, and measure cellular sensitivity to azathioprine using viability assays. Additionally, nucleotide analog incorporation assays and ROS detection provide complementary insights into the functional consequences of ITPA loss. For additional technical details or ordering information, please contact Ascent Research.
