The GNPAT Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HAP1 cells with targeted disruption of the GNPAT gene. This mixed population contains diverse gene-edited variants, each with unique CRISPR/Cas9-induced mutations at the GNPAT locus, collectively resulting in loss of functional gene expression. The polyclonal format avoids clonal selection artifacts and provides a heterogeneous knockout model suitable for assessing variable phenotypic responses.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myelogenous leukemia (CML) line. Its near-haploid karyotype facilitates CRISPR/Cas9-mediated gene disruption by reducing genetic redundancy, as most loci exist in a single copy. HAP1 cells are suspension-adapted and retain hematopoietic features, making them a valuable model for cancer biology, hematopoiesis, and signaling studies. The near-haploid background enhances knockout generation efficiency, though the polyclonal nature may include a low frequency of unedited cells.
GNPAT encodes peroxisomal dihydroxyacetone phosphate acyltransferase (DHAPAT), which catalyzes the first step in ether phospholipid biosynthesis: acylation of DHAP to 1-acyl-DHAP. This reaction is essential for subsequent processing by AGPS to produce plasmalogens and platelet-activating factor (PAF). GNPAT expression is regulated by PPARA and PPARG transcription factors, and its peroxisomal localization depends on interaction with the PEX5 receptor. Through these molecular associations, GNPAT controls membrane lipid composition and cellular antioxidant capacity.
In the HAP1 CML context, GNPAT knockout disrupts ether phospholipid synthesis, offering a model to explore the role of plasmalogens in hematopoietic cancer biology. Hematopoietic cells are particularly rich in plasmalogens, which contribute to membrane fluidity, trafficking, and oxidative stress resistance. Loss of GNPAT function impairs these processes and may affect cell proliferation and differentiation. This polyclonal knockout pool also enables modeling of rhizomelic chondrodysplasia punctata type 2 and peroxisomal disorders.
This product is suitable for lipidomic profiling of plasmalogen species, DHAPAT activity assays, and GNPAT immunodetection. Peroxisomal assessment by catalase staining or PEX5 immunofluorescence, and peroxisomal import flow cytometry using PTS1-tagged GFP, are applicable. Functional studies under oxidative stress, cell proliferation assays, and drug screening targeting ether lipid metabolism in cancer are representative applications. For further details, please contact Ascent Research.
