BCAM Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted disruption of the BCAM gene in the HAP1 human cell line. This polyclonal pool provides a loss-of-function model for investigating BCAM-dependent cellular processes, including cell adhesion, migration, and signal transduction. The knockout product format as polyclonal cells offers a heterogeneous population with diverse editing events, enabling robust functional analyses without clonal selection artifacts.
The HAP1 cell line is a near-haploid human cell model originally derived from the KBM-7 chronic myeloid leukemia (CML) cell line. Its near-haploid karyotype simplifies genetic manipulation and reduces gene redundancy, making it an ideal host for CRISPR/Cas9-mediated gene disruption. HAP1 cells are widely used in functional genomics and drug target validation due to their ease of culture and responsiveness to genetic perturbations. The CML origin also provides a cancer-relevant context for studying oncogenic signaling pathways.
BCAM encodes the Lutheran blood group glycoprotein (CD239), which functions as a receptor for laminin ??5 (LAMA5)-containing isoforms, particularly laminin-511 and -521. Upon ligand binding, BCAM activates focal adhesion kinase (FAK, PTK2) and Src family kinases, initiating downstream signaling through the PI3K/Akt and MAPK/ERK cascades. Key pathway components include PIK3CA, AKT1, and MAPK1 (ERK2). BCAM activity is regulated by transcription factors such as HIF1A, GATA1, SP1, and the cytokine TNF, and it interacts with cytoskeletal and membrane proteins including SPTB, ANK1, and CD44. This signaling network promotes cytoskeletal reorganization, cell spreading, and directed migration.
In the HAP1 host cell background, BCAM knockout enables dissection of laminin-driven adhesion signaling with minimal genetic compensation. The near-haploid state enhances the penetrance of the gene disruption, facilitating unambiguous genotype-phenotype correlations. This model is particularly useful for studying cancer cell dissemination, as BCAM-mediated adhesion contributes to metastatic spread in various carcinomas and hematologic malignancies. The knockout cells can be employed to assess the dependence of migration and invasion on BCAM-driven FAK/Src activation.
BCAM Knockout HAP1 Polyclonal Cells are amenable to diverse experimental assays, including Western blotting to confirm BCAM protein loss, flow cytometry to verify absence of CD239 surface expression, and adhesion assays on laminin-511 substrates. Migration and invasion can be evaluated using wound healing or Transwell methods, and phospho-FAK immunoblotting provides readout of FAK activation. Immunofluorescence imaging of actin stress fibers allows visualization of cytoskeletal changes downstream of BCAM signaling. For additional inquiries or to request custom cell models, please contact Ascent Research.
