The ITGAV Knockout HAP1 Polyclonal Cells are a heterogenous population of HAP1 cells with CRISPR/Cas9-mediated disruption of the ITGAV gene. This polyclonal knockout pool provides a loss-of-function model for studying integrin alpha V biology without the need for clone isolation, enabling evaluation of ITGAV-dependent phenotypes in a near-haploid genetic background.
The host HAP1 cell line is a near-haploid human chronic myeloid leukemia derivative of KBM-7, featuring an adherent, fibroblast-like morphology. Its largely haploid karyotype (with a disomic region on chromosome 8) simplifies gene knockout studies, as targeting a single allele is generally sufficient to eliminate gene function. HAP1 cells are widely used in functional genomics and CRISPR screens due to their stable growth and efficient editing.
ITGAV encodes integrin subunit alpha V, which heterodimerizes with beta subunits (ITGB3, ITGB5, ITGB6, ITGB8) to bind RGD-containing matrix proteins such as vitronectin and fibronectin. Ligand engagement activates FAK and SRC, which phosphorylate ERK1/2 and AKT through MAPK and PI3K cascades. ITGAV-containing integrins also activate latent TGF-beta via force transmitted through talin, kindlin, and the actin cytoskeleton. Upstream regulators include TGF-beta, EGF, VEGF, TNF-alpha, and transcription factors SP1 and HIF1A; downstream targets include FAK, SRC, ERK1/2, AKT, JNK, and SMAD2/3.
In the haploid HAP1 background, polyclonal ITGAV knockout produces a uniform loss of function, facilitating clear phenotypic interpretation. This model enables dissection of integrin-dependent adhesion, migration, and TGF-beta activation without diploid compensatory effects, and is well suited for high-throughput screens of integrin-targeted compounds.
Applications include cell adhesion assays on vitronectin or fibronectin, Boyden chamber migration/invasion tests, ELISA-based TGF-beta activation assays, and Western blotting for phospho-FAK and phospho-AKT. Immunofluorescence of focal adhesions, flow cytometry for integrin alpha V, and global profiling by RNA-seq or phospho-signaling analysis are also feasible. These cells support research in cancer metastasis, fibrosis, osteoporosis, Glanzmann thrombasthenia, and inflammatory diseases. For more information, contact Ascent Research.
