The ATP13A3 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ATP13A3 gene in the HEK293T host cell line. This knockout model provides a valuable tool for investigating the biological functions of ATP13A3, a P5-type ATPase implicated in endolysosomal cation and polyamine transport. By targeting ATP13A3, researchers can study loss-of-function phenotypes in a physiologically relevant cellular context.
The HEK293T cell line is a transformed human embryonic kidney epithelial cell line that stably expresses the SV40 large T-antigen, facilitating episomal replication of plasmids containing the SV40 origin of replication. Derived from HEK293 cells, which were originally generated by transformation with adenovirus 5 DNA, HEK293T cells are widely employed for their high transfection efficiency and robust protein expression capabilities. Their epithelial origin and well-characterized genetic background make them suitable for functional genomics studies, including CRISPR-mediated gene editing.
ATP13A3 encodes a P5-type cation-transporting ATPase that localizes to endolysosomal membranes, where it actively transports polyamines and cations across the membrane. This transport activity is essential for maintaining lysosomal ionic and polyamine homeostasis, which in turn regulates critical processes such as iron storage and autophagy. ATP13A3 is transcriptionally regulated by TFEB, a master regulator of lysosomal biogenesis, and its function is linked to downstream lysosomal polyamine levels, lysosomal iron storage, and autophagic flux. The protein interacts with lysosomal membrane proteins and may functionally collaborate with the related transporter ATP13A2. Dysregulation of ATP13A3 disrupts these pathways, leading to impaired lysosomal degradation and altered iron metabolism.
In the HEK293T background, knockout of ATP13A3 offers a powerful model to dissect the molecular mechanisms underlying endolysosomal trafficking and cation homeostasis. Given its association with pulmonary arterial hypertension and neurodegenerative diseases, including Parkinson??s disease spectrum disorders, this cell model enables the investigation of disease-relevant cellular phenotypes. The loss of ATP13A3 function can be studied in conjunction with upstream regulators such as TFEB and downstream effectors like ferritin and LC3, providing insights into pathophysiological signaling networks.
The ATP13A3 Knockout HEK293T Polyclonal Cells are suitable for a wide array of experimental approaches, including western blotting to confirm protein depletion, RT-qPCR for transcript analysis, immunofluorescence to visualize lysosomal markers, and LysoTracker staining to assess lysosomal pH changes. Functional assays such as polyamine uptake, iron quantitation, and autophagy flux measurement via LC3 turnover can directly evaluate the impact of ATP13A3 loss on cellular processes. Cell viability assays further allow the assessment of cellular stress responses. For additional information or custom inquiries, please contact Ascent Research.
