The HAPLN3 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population established from the near-haploid human HAP1 cell line. This loss-of-function model disrupts the HAPLN3 gene, which encodes a hyaluronan and proteoglycan link protein critical for stabilizing supramolecular aggregates in the extracellular matrix (ECM). The polyclonal population contains a mixture of cells with diverse knockout alleles, enabling analysis of gene function at the population level without isolation of individual clones.
The HAP1 host line is a male, chronic myeloid leukemia (CML)-derived, near-haploid cell line with a karyotype of 23 chromosomes. This near-haploid state simplifies loss-of-function studies by minimizing genetic redundancy, making HAP1 cells a preferred platform for CRISPR-based genetic screens. Despite their hematopoietic origin, HAP1 cells express adhesion molecules and ECM components, rendering them suitable for investigating ECM-cell interactions, integrin signaling, and migration.
HAPLN3 functions as a link protein that physically connects hyaluronan to proteoglycans such as aggrecan and versican, forming ternary complexes that maintain ECM integrity. Its transcription is positively regulated by SOX9 and TGF-??, while mechanical loading also modulates its expression. HAPLN3 interacts with hyaluronan, aggrecan, versican, CD44, and TSG-6. Knockout of HAPLN3 disrupts these networks, leading to impaired ECM structure, reduced integrin ??1 activation, and defective cell adhesion and migration. This disruption thereby impacts hyaluronan metabolism and downstream signaling cascades dependent on ECM attachment.
Within the HAP1 near-haploid context, HAPLN3 knockout provides a clean genetic background to dissect its specific contributions to ECM assembly and cell behavior without allelic compensation. The model is particularly suited for high-throughput phenotypic screens and genetic interaction studies aimed at uncovering compensatory link proteins or novel components of hyaluronan-mediated signaling.
Research applications span extracellular matrix biology, osteoarthritis and cartilage degeneration models, cancer metastasis, and drug target validation. Representative assays include Western blotting and RT-qPCR for expression analysis, hyaluronan binding assays to probe ECM complex formation, cell adhesion and migration assays to assess functional outcomes, and immunofluorescence to visualize ECM architecture. For additional technical details, please contact Ascent Research.
