The ITGA2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout cell population in which the ITGA2 gene has been functionally disrupted in the HAP1 host cell line. This polyclonal product provides a heterogeneous yet validated loss-of-function model, eliminating integrin alpha-2 expression and enabling robust phenotypic characterization of ITGA2-dependent processes without clonal selection bias.
HAP1 is a near-haploid human myeloid leukemia cell line derived from a patient with chronic myeloid leukemia. It was originally isolated as an adherent subclone of the KBM-7 cell line. The near-haploid karyotype permits efficient single-allele gene disruption, yielding complete loss-of-function phenotypes without allelic compensation, and has established HAP1 as a standard platform for CRISPR-based functional screens and knockout validation studies.
ITGA2 encodes the alpha-2 subunit of integrins, which heterodimerizes exclusively with beta-1 integrin (ITGB1) to form the major cellular receptor for collagen and laminin. Engagement of this receptor by its ligands triggers autophosphorylation of focal adhesion kinase (FAK) and Src kinases, activating the PI3K-AKT and MAPK/ERK signaling cascades that govern cell survival, proliferation, and migration. ITGA2 expression is regulated by upstream cytokines and transcription factors including TGF-beta, TNF-alpha, IL-1, HIF1A, SP1, NF-kB, and AP-1. Downstream targets include ERK1/2, AKT, matrix metalloproteinases MMP2 and MMP9, Cyclin D1, and Rho-family GTPases RAC1, CDC42, and RhoA. Key adaptor and scaffold proteins such as talin, kindlin, vinculin, paxillin, and caveolin-1 couple the integrin adhesion complex to the actin cytoskeleton.
Knockout of ITGA2 in HAP1 cells abolishes alpha2beta1 integrin surface expression, preventing adhesion to collagen- and laminin-rich matrices and silencing downstream FAK-Src-AKT-ERK signal transduction. This disruption impairs focal adhesion assembly and cytoskeletal remodeling, leading to defective cell spreading, migration, and invasion. The haploid background ensures that the knockout phenotype is clear and unattenuated, making this model particularly useful for genetic interaction mapping and chemical suppressor screens that probe integrin function.
Research applications include detailed studies of integrin-mediated adhesion, directional migration, and invasion in cancer and fibrosis models, as well as platelet function and thrombosis research. Typical assays compatible with this model encompass Western blot analysis of ITGA2 and phosphorylated signaling intermediates (pFAK, pAKT), adhesion and spreading assays on collagen or laminin, Boyden chamber migration/invasion assays, flow cytometry for surface integrin expression, immunofluorescence imaging of focal adhesion components, and RNA-sequencing for transcriptional profiling. These cells are also amenable to drug sensitivity testing with integrin inhibitors, supporting the validation of anti-metastatic therapeutic candidates. For more information or to place an order, please contact Ascent Research.
