The ATP8B2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting ATP8B2 in the HAP1 human haploid cell line. This gene encodes a P4-ATPase phospholipid flippase. The polyclonal pool contains a variety of loss-of-function alleles, providing a robust model free from single-clone bias. This product is suitable for studying ATP8B2 deficiency in a range of functional assays.
HAP1 cells are a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia, exhibiting adherent fibroblast-like morphology. With only one copy of most chromosomes, a single gene disruption suffices to produce a complete knockout phenotype. This genetic simplicity makes HAP1 cells an ideal host for CRISPR/Cas9-mediated knockout studies, widely used in mutagenesis screens and phenotypic analyses.
ATP8B2 functions as a phosphatidylserine flippase, translocating phosphatidylserine from the exoplasmic to cytoplasmic membrane leaflet to maintain lipid asymmetry. It partners with TMEM30A (CDC50A) for proper activity. Knockout of ATP8B2 abolishes flippase activity, causing aberrant phosphatidylserine exposure that recruits ESCRT machinery, alters endolysosomal sorting, and affects downstream signaling like PKC activation. These disruptions impair vesicular trafficking and are linked to auditory hair cell maintenance.
In the HAP1 context, this knockout model enables dissection of ATP8B2’s role in membrane trafficking and signaling with clear genotype-phenotype correlations. The near-haploid karyotype avoids complications from compensating wild-type alleles. The model is valuable for investigating hearing loss pathologies such as DFNB107 and auditory neuropathy spectrum disorder, where ATP8B2 mutations are implicated.
Applications include western blotting for knockout confirmation, immunofluorescence for localization studies, and flow cytometry with Annexin V to measure phosphatidylserine exposure. Co-immunoprecipitation can assess ATP8B2-TMEM30A interaction. The cells are also suitable for drug screening targeting lipid transport disorders and phenotypic rescue experiments. For further information, contact Ascent Research.
