The B3GALT6 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid HAP1 cell line. This heterogeneous knockout pool abrogates B3GALT6 function across the population, enabling robust loss-of-function studies. The polyclonal format ensures comprehensive target disruption while maintaining biological representativeness, making it ideal for glycobiology, extracellular matrix biology, and disease modeling.
The HAP1 cell line is an adherent fibroblast-like cell line originating from the near-haploid KBM-7 chronic myelogenous leukemia line from a male donor. Its near-haploid karyotype??with single copies of most chromosomes??facilitates efficient CRISPR/Cas9-mediated gene knockout, as altering one allele often abolishes gene function. HAP1 cells are widely used in genetic screens and knockout studies due to their reproducible editing outcomes and human physiological relevance.
B3GALT6 encodes a galactosyltransferase that catalyzes the transfer of galactose from UDP-galactose to xylose, a critical step in the synthesis of the proteoglycan linkage tetrasaccharide. This enzyme acts downstream of XYLT1/XYLT2 and B4GALT7, and upstream of B3GAT3 and the EXT1/EXT2 elongation complex, forming the core region for heparan sulfate and chondroitin sulfate chain assembly. Disruption of B3GALT6 blocks glycosaminoglycan chain initiation, resulting in defective proteoglycan synthesis and altered extracellular matrix. Signaling pathways dependent on heparan sulfate proteoglycans, such as TGF-?? pathways, are thereby compromised.
In the near-haploid HAP1 context, B3GALT6 knockout allows unambiguous investigation of proteoglycan dysfunction without diploid gene compensation. This model is particularly suited for studying the molecular basis of Ehlers-Danlos syndrome (spondylodysplastic type) and spondyloepimetaphyseal dysplasia with joint laxity, both linked to B3GALT6 mutations. Researchers can use these cells to dissect how impaired glycosaminoglycan synthesis disrupts cell adhesion, migration, and extracellular matrix integrity.
These cells support a variety of assays, including RT-qPCR for B3GALT6 transcript, western blotting for proteoglycan cores (syndecan, glypican), immunofluorescence for heparan sulfate, and GAG disaccharide analysis by mass spectrometry. Functional assays such as cell migration, ECM staining, and flow cytometry for surface proteoglycans assess biological outcomes. For technical inquiries, please contact Ascent Research.
