The ITGA5 Knockout HAP1 Polyclonal Cells are a polyclonal population of HAP1 cells with CRISPR/Cas9-mediated disruption of the ITGA5 gene. This loss-of-function model enables the study of integrin alpha-5 in a heterogeneous knockout context, avoiding the limitations of clonal selection. The polyclonal format provides a diverse genetic background suitable for pooled screening applications.
HAP1 is a near-haploid human cell line derived from a male chronic myeloid leukemia patient. It contains the BCR-ABL fusion and grows in suspension, making it ideal for high-throughput genetic and biochemical studies. The near-haploid genome simplifies gene targeting and reduces functional redundancy, offering a clean background for investigating signaling pathways.
ITGA5 encodes the integrin alpha-5 subunit, which pairs with ITGB1 to form the fibronectin receptor. Ligand binding triggers recruitment and activation of FAK (PTK2) and SRC, leading to downstream signaling through AKT1 and MAPK1/3 (ERK1/2). Key adaptors include TLN1, FERMT2, VCL, and PXN, linking the receptor to the actin cytoskeleton. Upstream regulators such as TGFB1 and EGF control ITGA5 expression, while downstream pathways regulate cell adhesion, migration, proliferation, and survival.
Disruption of ITGA5 in HAP1 cells provides a clean system for dissecting integrin-dependent phenotypes. The near-haploid background minimizes gene dosage complications, and the loss of alpha-5 integrin impairs fibronectin adhesion, directly affecting downstream signaling. This model is particularly useful for investigating cell-ECM interactions and identifying genetic interactions in pooled screens.
Applications include cancer biology studies of adhesion and migration, drug target validation, and functional genomics. Typical assays are flow cytometry for alpha-5 surface expression, fibronectin adhesion assays, FAK phosphorylation Western blots, and migration/invasion assays. Co-immunoprecipitation can probe integrin complex integrity. For more information, please contact Ascent Research.
