ITFG2 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the ITFG2 gene has been disrupted to ablate expression of the integrin alpha FG-GAP repeat containing 2 (ITFG2) protein. This loss-of-function model is generated via CRISPR/Cas9-mediated gene disruption in the near-haploid HAP1 cell line, yielding a heterogeneous pool of edited cells that reflects a population-level knockout. Supplied as ready-to-use polyclonal cells, this product enables direct investigation of ITFG2-dependent processes in integrin signaling and cell adhesion without the clonal selection bias inherent to monoclonal lines.
HAP1 cells are a near-haploid human cell line derived from the chronic myeloid leukemia KBM-7 line. Their haploid karyotype provides a simplified genetic background that facilitates efficient genome editing and unambiguous assignment of genotype to phenotype, making them a premier model for functional genomics screens, knockout validation, and pathway dissection. Despite their haploid nature, HAP1 cells maintain robust expression of adhesion receptors and focal adhesion components, rendering them a physiologically relevant platform for dissecting integrin-mediated signaling and cytoskeletal dynamics.
ITFG2 is an integrin adaptor protein that localizes to focal adhesions, where it directly binds integrin beta 1 (ITGB1) and talin (TLN1). This interaction couples integrin receptors to the actin cytoskeleton, forming a critical link in the adhesion complex. Following integrin activation by extracellular matrix ligands and mechanical force, ITFG2 facilitates the recruitment and autophosphorylation of focal adhesion kinase (PTK2/FAK) at Tyr397. FAK, in turn, activates downstream effectors including Src kinase, paxillin, and vinculin, which promote actin polymerization and focal adhesion turnover. Additional interacting partners such as kindlin contribute to the maturation of adhesion sites. Disruption of ITFG2 abolishes this scaffolding function, leading to attenuated FAK signaling, impaired cytoskeletal reorganization, and defective cell adhesion and migration.
In the HAP1 haploid background, the ITFG2 knockout model offers a clean genetic system to interrogate focal adhesion signaling with minimal compensatory complexity. The absence of a second allele ensures that any observed phenotypic alterations can be directly attributed to ITFG2 deficiency, enhancing the interpretability of mechanistic studies. This model is particularly well-suited for high-resolution live-cell imaging of adhesion dynamics and for quantitatively assessing migration and adhesion parameters. The polyclonal nature of the population preserves editing diversity, making it compatible with pooled screening strategies while still supporting robust biochemical and cell biological assays.
Key research applications include Western blotting for phosphorylated FAK (pTyr397) and Src substrates to monitor pathway activation, co-immunoprecipitation to map ITFG2-containing protein complexes, and immunofluorescence microscopy of focal adhesion markers such as paxillin and vinculin. Functional assays, including adhesion to fibronectin- or collagen-coated surfaces and transwell migration tests, enable quantitative evaluation of cellular phenotypes. Additionally, this knockout pool serves as a powerful tool for haploid genetic screens aimed at identifying synthetic lethal partners or novel regulators of integrin signaling. For product inquiries and technical support, please contact Ascent Research.
