The B3GALNT2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid HAP1 human cell line. This product provides a loss-of-function model for the B3GALNT2 gene, which encodes a critical beta-1,3-N-acetylgalactosaminyltransferase. The polyclonal pool is generated by CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous cell population lacking functional B3GALNT2 protein. This model is designed for researchers investigating LacdiNAc glycosylation in cell adhesion, signaling, and disease.
HAP1 is a near-haploid human cell line originally derived from the BCR-ABL-positive chronic myeloid leukemia KBM-7 line. Its stable haploid karyotype enables straightforward generation of recessive knockout models and is well suited for genetic screening and gene function studies. The near-haploid background eliminates gene dosage complexity, making HAP1 an optimal host for studying complete gene disruption phenotypes in a simplified genomic context.
B3GALNT2 functions as a glycosyltransferase that adds GalNAc to terminal GlcNAc residues on dystroglycan and integrins, forming LacdiNAc structures essential for cell adhesion and basement membrane organization. This activity is integrated within the dystroglycan glycosylation pathway, which includes POMT1, POMT2, POMGNT1, B3GALNT2, and LARGE. B3GALNT2 is transcriptionally regulated by SP1 (predicted) and responds to developmental cues. Its disruption impairs dystroglycan glycosylation, laminin binding, and integrin ??3??1-mediated adhesion, leading to defective cell-ECM interactions.
In HAP1 cells, B3GALNT2 knockout abolishes LacdiNAc synthesis, enabling dissection of glycosylation-dependent adhesion and signaling. The near-haploid background simplifies analysis of complete gene loss, making this model particularly relevant for dystroglycanopathy diseases such as congenital muscular dystrophy-dystroglycanopathy type A11 (Walker-Warburg syndrome) and for studying cancer metastasis, where altered glycosylation drives invasive behavior.
Researchers can employ this knockout population in functional assays such as western blotting with anti-B3GALNT2 and WFA lectin, immunofluorescence for dystroglycan and integrin localization, cell adhesion and migration assays, flow cytometry for LacdiNAc, RT-qPCR, and mass spectrometry-based glycan profiling. These tools facilitate studies of glycosylation in metastasis, dystroglycanopathy disease modeling, haploid genetic screens for glycosylation phenotypes, and drug target validation for muscular dystrophy. For further information, contact the Ascent Research support team.
