The ASAH1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the ASAH1 gene in the HAP1 cell line. This product delivers a loss-of-function model for studying acid ceramidase, enabling analysis of sphingolipid metabolism and its downstream effects in a near-haploid genetic background without clonal selection.
HAP1 is a fibroblast-like, adherent human near-haploid cell line derived from CML line KBM-7. Its near-haploid karyotype simplifies functional genomics by reducing genetic redundancy, enhancing knockout efficiency and phenotype interpretation. The cells retain key signaling and metabolic pathways relevant to cancer and lysosomal biology, providing a disease-relevant platform for ASAH1 knockout studies.
ASAH1 encodes lysosomal acid ceramidase, which hydrolyzes ceramide to sphingosine and fatty acid, a critical step in the ceramide-to-sphingosine-1-phosphate (S1P) conversion. Regulated by TNF-??, IL-1??, TFEB, and lysosomal pH, acid ceramidase interacts with Saposin D and Prosaposin for activity and is influenced by cathepsin A. By reducing pro-apoptotic ceramide and generating sphingosine for S1P synthesis via sphingosine kinases, ASAH1 promotes cell survival and attenuates ceramide-mediated apoptosis. This places ASAH1 within a network of ceramide synthases, sphingomyelinases, ceramidases, sphingosine kinases, S1P phosphatases, S1P lyase, and S1P receptors.
Disrupting ASAH1 in HAP1 cells uncouples the ceramide/S1P rheostat, providing a clear model for studying apoptosis resistance, lysosomal degradation defects, and sphingolipid imbalance. The near-haploid state ensures unambiguous genotype-phenotype linkage, making it ideal for investigating mechanisms underlying Farber lipogranulomatosis and cancer drug resistance, where ASAH1 overexpression is implicated.
These cells support diverse assays such as Western blotting, RT-qPCR, lipid mass spectrometry, apoptosis and viability assays, and immunofluorescence. Applications include cancer apoptosis resistance studies, lysosomal storage disease modeling, and drug resistance screening. For additional information, contact Ascent Research.
