This product consists of a CRISPR/Cas9-edited polyclonal knockout population of HAP1 cells with disruption of the HMMR gene (RHAMM). The polyclonal format provides a heterogeneous pool of edited alleles, establishing a loss-of-function model without clonal selection. The HMMR-targeting disruption ablates functional protein expression, enabling robust investigation of hyaluronan-mediated motility, spindle regulation, and associated signaling in a near-haploid background.
HAP1 is a near-haploid human fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia line (male origin). Its near-haploidy facilitates genetic analysis by minimizing gene redundancy, making it an ideal platform for knockout studies requiring clear genotype-phenotype relationships. HAP1 cells retain responsiveness to growth factors and extracellular matrix cues, supporting detailed studies of adhesion, migration, and cytoskeletal dynamics.
HMMR functions as both a cell-surface hyaluronan receptor and an intracellular microtubule-associated protein. Activated by EGF, TGF-??, and hypoxia, HMMR drives cell motility and proliferation mainly via ERK1/2 and FAK signaling, leading to upregulation of MMP-9 and Cyclin D1. It forms co-receptor complexes with CD44 at the membrane and interacts with dynein, BRCA1, and tubulin to regulate mitotic spindle integrity. Key downstream mediators include SRC and Rho GTPases, integrating focal adhesion dynamics and cell cycle progression.
In the HAP1 context, HMMR disruption permits precise dissection of its roles without allelic buffering, unmasking phenotypes in spindle assembly, migration, or cell cycle progression. The fibroblast-like morphology makes these cells particularly suited for studying mesenchymal motility and adhesion relevant to metastasis and wound healing. This model helps elucidate how HMMR loss alters the crosstalk between hyaluronan signaling, FAK/ERK activation, and microtubule organization.
The HMMR-knockout polyclonal HAP1 cells are suitable for Transwell migration and wound healing assays to assess motility deficits, immunofluorescence for mitotic spindle analysis, flow cytometry for cell cycle profiling, and Western blotting for ERK/FAK phosphorylation. Co-immunoprecipitation can probe disrupted CD44 or dynein interactions. The model supports drug target validation for cancers (breast, gastric, prostate) and hyaluronan-related inflammation such as rheumatoid arthritis. For additional technical information, please contact Ascent Research.
