The ATP13A2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HAP1 human near-haploid cell line, providing a loss-of-function model for ATP13A2. This gene encodes a lysosomal P5-type ATPase transporter; its disruption enables detailed investigation of lysosomal biology, autophagy, and neurodegenerative disease pathways.
HAP1 is a near-haploid human suspension cell line derived from the KBM-7 chronic myeloid leukemia line. Its haploid genome simplifies genetic knockout because a single targeting event can disrupt gene function, and p53 deficiency facilitates efficient genome editing. Grown in suspension, HAP1 cells are well-suited for scalable biochemical assays and high-throughput screening, making them a favored platform for CRISPR-based functional genomics studies.
ATP13A2 functions as a lysosomal P5-type ATPase transporter that mediates polyamine and cation export, essential for lysosomal pH homeostasis and proteolytic capacity. Its expression is transcriptionally regulated by TFEB, HIF1A, and NRF2 in response to mTORC1 inhibition and lysosomal stress. Loss of ATP13A2 leads to lysosomal alkalinization, impaired autophagosome-lysosome fusion, and reduced cathepsin D activity, causing accumulation of autophagic substrates p62/SQSTM1 and LC3-II. ATP13A2 interacts with LAMP2, ??-synuclein, HDAC6, HSP70, and CLN5. Its deficiency disrupts ??-synuclein clearance and mitochondrial quality control, affecting mitophagy regulators MFN1/2 and the iron transporter DMT1, thereby linking lysosomal dysfunction to Parkinson’s disease pathogenesis.
Within the HAP1 near-haploid background, disruption of a single ATP13A2 allele generates a clear loss-of-function phenotype without allelic compensation, thereby increasing the penetrance of autophagy and lysosomal defects. Although HAP1 cells originate from a leukemic lineage, they retain the core autophagy-lysosomal machinery, allowing investigation of neurodegeneration-relevant mechanisms in a human cell system. The p53 deficiency simplifies genome editing and, for most ATP13A2-related processes, does not confound the interpretation of lysosomal and mitochondrial phenotypes. This genetic context makes the polyclonal knockout population a robust and accessible model.
This knockout model supports diverse research applications, including Parkinson’s disease modeling, autophagy flux analysis (Western blotting for LC3-II and p62), lysosomal integrity assessment (immunofluorescence for LAMP1 and cathepsin D, LysoSensor pH measurement), and ??-synuclein aggregation assays. Additional uses encompass mitochondrial morphology evaluation with MitoTracker, iron accumulation detection with FerroOrange, and high-throughput screening for TFEB-activating compounds. Polyamine transport studies and investigation of ATP13A2?CLN5 interactions in neuronal ceroid lipofuscinosis are also enabled. For detailed technical information or custom knockout cell inquiries, please contact Ascent Research.
