B4GALT6 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the B4GALT6 gene in the HAP1 cell line. This heterogeneous pool is generated via CRISPR/Cas9-mediated gene disruption, minimizing clonal selection bias inherent in monoclonal lines. The polyclonal format enables robust loss-of-function studies of glycosphingolipid biosynthesis while preserving genetic complexity. These cells are suitable for investigating the functional consequences of B4GALT6 ablation in a haploid, chronic myeloid leukemia-derived background.
HAP1 cells are an adherent, fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia (CML) cell line. These near-haploid cells offer a simplified genetic landscape, facilitating knockout studies without the complication of diploid gene compensation. As a CML model, HAP1 cells retain oncogenic signaling features relevant to leukemia research while also providing a generic platform for exploring fundamental cell biology. The B4GALT6 knockout in this background is particularly useful for studying glycosphingolipid alterations in the context of hematopoietic malignancies.
B4GALT6 encodes a beta-1,4-galactosyltransferase that catalyzes the transfer of galactose from UDP-galactose to glucosylceramide (GlcCer) to form lactosylceramide (LacCer). This reaction is a pivotal step in the biosynthesis of complex glycosphingolipids, including gangliosides and globosides. B4GALT6 is transcriptionally regulated by SP1 and TFAP2A and interacts with glucosylceramide synthase and ceramide glucosyltransferase in the Golgi. Pathway components such as ceramide and GM3 synthase operate upstream and downstream, respectively, positioning B4GALT6 at a critical branch point. Disruption of B4GALT6 interrupts the production of LacCer and its downstream derivatives, altering membrane microdomain organization and sphingolipid-dependent signaling cascades.
In the HAP1 CML context, B4GALT6 knockout holds significant relevance for cancer glycobiology. Aberrant glycosylation is a hallmark of cancer, and altered glycosphingolipid expression contributes to malignant phenotypes including proliferation, migration, and drug resistance. By eliminating B4GALT6 activity, researchers can dissect the specific roles of lactosylceramide and its downstream metabolites in leukemogenesis and tumor microenvironment interactions. Moreover, this model serves as a platform for investigating congenital disorders of glycosylation linked to B4GALT6 dysfunction and for exploring Gaucher disease-related sphingolipid perturbations.
Typical applications include profiling glycosphingolipid changes via mass spectrometry lipidomics, measuring lactosylceramide synthase activity, and performing lectin-based flow cytometry to detect surface glycosylation changes. The knockout cells can be assayed for proliferation and migration alterations, providing functional readouts of sphingolipid-dependent signaling. Researchers may also use this model to screen compounds that modulate glycosphingolipid pathways or to study drug resistance mechanisms involving membrane lipid remodeling. For further details or to inquire about this product, please contact Ascent Research.
