The ATP13A3 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, featuring targeted disruption of the ATP13A3 gene. This pooled format provides an isogenic background for studying ATP13A3 deficiency without clonal bias, enabling functional analysis in a heterogeneous population. The product is supplied as ready-to-use polyclonal cells generated via CRISPR/Cas9-mediated gene disruption, suitable for applications requiring abrogation of ATP13A3 expression. The knockout relies on general Cas9-mediated target-gene disruption to impair protein function.
The HeLa host cell line is an HPV18-positive immortalized cervical adenocarcinoma line, widely used in cancer research for its robust growth and well-characterized signaling networks. HeLa cells are well-suited for studying polyamine metabolism and mTORC1 signaling due to their high proliferation and frequent use in mTOR pathway analysis. The HeLa background allows direct investigation of ATP13A3-mediated processes in a cancer context, where polyamine homeostasis and mTORC1 activity are often dysregulated.
ATP13A3 encodes a P5B-type ATPase that functions as an endosomal polyamine transporter, primarily shuttling spermidine from the endosomal lumen into the cytosol. This activity maintains polyamine homeostasis and modulates mTORC1 signaling. Upstream regulators such as c-MYC and HIF1A, along with cellular polyamine levels, control ATP13A3 expression and activity. Spermidine delivery activates mTORC1, phosphorylating S6K and 4E-BP1 to promote protein synthesis and inhibit autophagy. Polyamine metabolic enzymes (ODC1, SRM, SMS, PAOX, SMOX) coordinate with ATP13A3 to regulate polyamine flux. Consequently, ATP13A3 acts as a critical node linking endocytic polyamine import to nutrient-sensing and growth control.
In the HeLa cell model, disruption of ATP13A3 perturbs polyamine transport, reducing cytosolic spermidine and impairing mTORC1 activation. This mimics conditions in pulmonary arterial hypertension (PAH) and cancers with altered polyamine metabolism. HeLa cells exhibit high mTORC1 activity and polyamine dependence, making them sensitive to ATP13A3 loss. Thus, this model is valuable for studying polyamine-dependent mTORC1 signaling in tumor growth and autophagy.
This product is suited for studying ATP13A3-mediated polyamine transport, mTORC1 signaling, and drug target validation. Assays include western blotting for p-S6K/p-4E-BP1, LC-MS polyamine quantification, MTT proliferation assay, LC3-II autophagy flux measurement, and immunofluorescence for ATP13A3. Rescue experiments with exogenous spermidine confirm phenotype specificity. For detailed technical specifications, please contact Ascent Research.
