The ASRGL1 Knockout SK-HEP-1 Polyclonal Cells product comprises a heterogeneous population of SK-HEP-1 cells genetically modified using CRISPR/Cas9 technology to achieve targeted disruption of the ASRGL1 gene. These polyclonal knockout cells are supplied as a mixed population, reflecting a range of editing events across the cell pool, which is advantageous for studying gene function in a biologically variable context without relying on single-clonal artifacts. This product is designed for advanced biomedical research, particularly in cancer biology, amino acid metabolism, and tumor suppressor analysis.
The parental SK-HEP-1 cell line is derived from the ascitic fluid of a male patient with liver adenocarcinoma and displays endothelial-like characteristics, making it a unique model for hepatocellular carcinoma. These cells exhibit features relevant to tumor microenvironment interactions and metastatic progression. The endothelial-like phenotype and hepatic origin provide a distinctive platform for examining ASRGL1 function in liver cancer, where metabolic reprogramming and angiogenesis are critical.
ASRGL1 encodes an enzyme with dual asparaginase and isoaspartyl peptidase activities, catalyzing the hydrolysis of L-asparagine to L-aspartate and ammonia. As a putative tumor suppressor, ASRGL1 limits intracellular asparagine availability, thereby suppressing mTORC1 signaling and cell proliferation. In this knockout model, loss of ASRGL1 function leads to elevated asparagine levels, enhanced activation of the mTORC1 complex, and downstream phosphorylation of effectors such as p70S6K and 4E-BP1, while modulating p21 and Bax expression. Upstream regulators include p53 and ATF4, which integrate nutrient deprivation signals. The pathway also involves ASNS and Raptor, with engagement of Caspase-3 in apoptotic responses. ASRGL1 forms a functional homodimer and processes beta-aspartyl dipeptides, linking amino acid metabolism to cell growth control.
In the context of SK-HEP-1 cells, ASRGL1 knockout accentuates oncogenic traits by derepressing mTORC1 activity and promoting survival and proliferation. The endothelial-like properties of this cell line offer a model for investigating how asparagine homeostasis influences tumor angiogenesis and invasion. Because ASRGL1 is often downregulated in liver adenocarcinoma, this polyclonal knockout population provides a relevant system to study its tumor-suppressive mechanisms and to evaluate metabolic therapies that target asparagine dependency. Researchers can use this model to dissect the interplay between ASRGL1 and other tumor suppressors or oncogenes in a liver cancer background.
This polyclonal knockout cell product supports a wide array of experimental approaches, including Western blotting to assess ASRGL1, phosphorylated mTOR, and p70S6K levels, RT-qPCR for transcript analysis, and LC-MS-based quantification of asparagine and glutamine to probe metabolic changes. Functional assays such as MTT proliferation, Annexin V/PI apoptosis, colony formation, and soft agar growth can delineate the knockout phenotype in vitro. Furthermore, xenograft tumor models enable in vivo evaluation of tumor growth and therapeutic responses. These cells are suitable for drug discovery efforts seeking ASRGL1 mimetics or mTORC1 inhibitors, as well as for fundamental studies on nutrient-sensing and tumor suppression. For professional inquiries or technical support, please contact Ascent Research.
