The ITGB4 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ITGB4 gene in the near-haploid HAP1 cell line. This heterogeneous pool of edited cells avoids clonal bias and enables population-level functional studies of integrin ??4.
HAP1 is a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia cells. With its mostly haploid genome, HAP1 provides a simplified genetic background that facilitates knockout generation and eliminates concerns about a second functional allele. The adherent cells proliferate robustly and retain key adhesion and signaling pathways, making them a valuable platform for genetic screening and signaling studies.
ITGB4 encodes integrin ??4, which dimerizes with ITGA6 to form the laminin-332 receptor ??6??4. In epithelial cells, ??6??4 is a core component of hemidesmosomes, anchoring intermediate filaments via plectin, BP180, and BP230. Upon ligand binding, the integrin recruits Shc and Grb2, activating PI3K-Akt and MAPK/ERK cascades that phosphorylate AKT1 and MAPK1 (ERK2) and regulate FAK and Rac1. ITGB4 signaling is modulated by upstream factors like EGF, TGF-??1, HGF, and mechanical stress, positioning it as a hub for adhesion and growth factor crosstalk.
In the HAP1 model, ITGB4 disruption abolishes laminin-dependent adhesion and downstream signal transduction. Although HAP1 cells are leukemic rather than epithelial, they express ??6??4-interacting partners, enabling mechanistic dissection of ITGB4 function without epithelial-specific complexity. The haploid state enhances genotype-phenotype correlation, making this model ideal for studying hemidesmosome biology, integrin signaling, and the molecular consequences of ITGB4 loss.
This polyclonal knockout population is suited for diverse applications, including Western blotting and RT-qPCR for knockout validation, immunofluorescence for hemidesmosome components, adhesion and migration assays on laminin, and flow cytometry for surface integrin expression. Transcriptomic profiling by RNA-seq and phospho-AKT/ERK analysis can reveal downstream pathway alterations. These tools support research into epidermolysis bullosa, cancer metastasis, EMT, and adhesion-related drug discovery. For additional information, please contact Ascent Research.
