The B3GAT3 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population for targeted disruption of the B3GAT3 gene. This product comprises a heterogeneous pool of HAP1 cells harboring diverse loss-of-function mutations, generated without single-cell cloning to avoid clonal bias. The polyclonal format is advantageous for population-level functional studies and avoids artifacts of monoclonal selection.
HAP1 is a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia cells, engineered for adherent growth and stable haploidy. Its single-copy genome permits efficient gene disruption with a single targeting event, making it ideal for knockout and haploid genetic screening. HAP1 cells maintain essential mammalian signaling pathways and are extensively used for reverse genetics and biosynthetic pathway analysis.
B3GAT3 encodes glucuronyltransferase I, which catalyzes the addition of glucuronic acid to the proteoglycan linker tetrasaccharide, a prerequisite for chondroitin sulfate and heparan sulfate chain elongation. The enzyme functions downstream of xylosyltransferase and galactosyltransferases and is transcriptionally regulated by SOX9 and TGF-beta signaling. Its activity enables the subsequent action of chondroitin sulfate and heparan sulfate synthases on core proteins such as aggrecan, versican, perlecan, and syndecans. Knockout of B3GAT3 halts glycosaminoglycan biosynthesis, leading to aberrant proteoglycan maturation and disrupted extracellular matrix signaling.
In the HAP1 haploid model, B3GAT3 disruption creates a robust loss-of-function system to study early glycosaminoglycan assembly. The polyclonal population supports pooled screening approaches and functional genomics studies investigating proteoglycan roles in cell adhesion, migration, and matrix organization. This model is especially relevant for connective tissue disease research, including spondyloepimetaphyseal dysplasia with joint laxity and Larsen syndrome, as well as cancer and aortic aneurysm pathologies.
Research applications include proteoglycan biosynthesis studies, extracellular matrix assembly assays, and cancer cell migration analyses. Representative techniques comprise Western blotting for proteoglycan core proteins, immunofluorescence detection of glycosaminoglycan chains, and mass spectrometry-based disaccharide profiling. Functional readouts such as Alcian blue staining, flow cytometry for cell surface heparan sulfate, and cell adhesion/migration assays can delineate phenotypic consequences of B3GAT3 loss. For detailed information and technical support, please contact Ascent Research.
