The AS3MT Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population with disruption of the AS3MT gene in the HeLa human cervical adenocarcinoma cell line. This knockout model is provided as a heterogeneous pool, avoiding clonal selection, and is designed for functional studies of arsenic metabolism. The polyclonal format captures a range of editing events, making it suitable for population-level analyses of gene function.
HeLa cells are an immortalized epithelial cell line derived from a cervical adenocarcinoma, widely employed in cancer research, toxicology, and drug metabolism. Their rapid proliferation, ease of handling, and well-characterized signaling pathways allow for reproducible investigations into cellular responses to xenobiotics, including arsenic compounds.
AS3MT (arsenic methyltransferase) is the key enzyme in the biomethylation of inorganic arsenite (AsIII) to monomethylarsonate (MMA) and dimethylarsenate (DMA), utilizing S-adenosylmethionine (SAM) as the methyl donor. This detoxification pathway is essential for converting highly toxic trivalent arsenic into less reactive species that are effluxed by multidrug resistance-associated proteins (MRPs). AS3MT activity is regulated by the transcription factor NRF2, which is induced by oxidative stress and arsenic exposure, linking detoxification to cellular antioxidant defenses. The enzyme??s function is supported by reducing cofactors such as glutathione, thioredoxin, and glutaredoxin, which maintain its catalytic cysteine residues in a reduced state. Disruption of AS3MT abolishes this methylation capacity, resulting in intracellular accumulation of arsenite and increased susceptibility to arsenic-induced oxidative damage and cytotoxicity.
In HeLa cells, AS3MT knockout provides a valuable tool for studying arsenic biotransformation in a cancer-relevant epithelial context. HeLa cells natively express aquaglyceroporin AQP9, which mediates arsenite uptake, and harbor p53 inactivation, a common feature in cancers associated with arsenic exposure. This genetic background allows researchers to explore how AS3MT deficiency influences arsenic-induced DNA damage, apoptosis, and long-term carcinogenic outcomes. The model can be used to dissect the interplay between one-carbon metabolism and redox buffering in the setting of impaired methylation.
Researchers can employ this polyclonal knockout population in a variety of assays, including western blotting and RT-qPCR for confirming AS3MT ablation, HPLC-ICP-MS for quantitative arsenic speciation, methylation activity assays, and cellular uptake/efflux experiments. The product is well-suited for toxicological assessments of arsenic-related skin lesions, bladder and lung cancers, and cardiovascular disease. For additional information, please contact Ascent Research.
